Shotgun immunoproteomics to identify diseaseassociated bacterial antigens:: Application to human colon cancer

Circulating antibodies reflect a mirror view of invading antigens that are related to infection and cancer. This was recently exemplified by using serum antibodies to capture Streptococcus bovis antigens followed by M S to generate antigen profiles that were diagnostic for colon cancer. These bacterial antigen profiles have a high potential to aid in the immuno-diagnosis of this disease, as the magnitude of the immune response to bacterial antigens is, in general, superior to the immune response against tumor (self) antigens. In this study, the identity of individual colon cancer-associated streptococcal antigens was revealed by enrichment of these "diagnostic" antigens by selected patient antibodies followed by high-accuracy nanoLC-MS/MS peptide identification. This showed that both the histone-like protein HlpA and the ribosomal protein Rp L7/L12 are members of the colon cancer-associated S. bovis immunome. Both antigens also seem to belong to the group of anchorless surface proteins, like 14 additional proteins that were co-identified in S. bovis cell wall extracts. Among these were the known streptococcal anchorless surface proteins GAPDH and Enolase. Taken together, these data show that shotgun immunoproteomics, combining immunocapture in-line with LC MS/MS, is a convenient approach for the rapid identification of disease-associated bacterial antigens.