Large T antigen variants of human polyomaviruses 9 and 12 and seroreactivity against their N terminus

The tumour antigens (TAgs) of mammalian polyomaviruses (PyVs) are key proteins responsible for modulating the host cell cycle and are involved in virus replication as well as cell transformation and tumour formation. Here we aimed to identify mRNA sequences of known and novel TAgs encoded by the recently discovered human polyomaviruses 9 and 12 (HPyV9 and HPyV12) in cell culture. Synthetic viral genomes were transfected into human and animal cell lines. Gene expression occurred in most cell lines, as measured by quantitative PCR of cDNA copies of mRNA encoding major structural protein VP1. Large TAg- and small TAg-encoding mRNAs were detected in all cell lines, and additional spliced mRNAs were identified encoding TAg variants of 145 aa (HPyV9) and 84 aa (HPyV12). Using as antigens in ELISA the N-terminal 78 aa common to all respective TAg variants of HPyV9 and HPyV12, seroreactivity of 100 healthy blood donors, 54 patients with malignant diseases of the gastrointestinal tract (GIT) and 32 patients with non-malignant diseases of the GIT was analysed. For comparison, the corresponding TAg N termini of BK PyV (BKPyV) and Merkel cell PyV (MCPyV) were included. Frequent reactivity against HPyV9, HPyV12 and BKPyV TAgs, but not MCPyV TAg, was observed in all tested groups. This indicates expression activity of the early region of three human PyVs in healthy and diseased subjects.