The human polyomavirus, JCV, causes a demyelinating disease in immune compromised individuals, progressive multifocal leukoencephalopathy (PML). PML is a substantial neurological complication in AIDS patients, occuring in 8% of all cases. PML is the AIDS defining illness in almost 1% of AIDS cases. Although in the past PML was a rare disease, its incidence is high in immune compromised patients like AIDS and allograft recipients. JC Virus was orignially thought to be strictly neurotropic, infecting only macroglial cells derived from the human brain. However, our results have now shown that there is a firm link between susceptible lymphoid and glial cells in the pathogenesis of JCV infection leading to demyelination in the human brain. The data indicate that a similar viral genotype which is found in brain tissue of PML patients originates from an initial lymphoid cell infection, including the bone marrow. The molecular similarities for JCV gene expression between cells of the nervous and immune systems directly correlated with the expression of the NF-1 class X family of DNA binding proteins which function for viral transcription. We have cloned the gene for NF-1/X from human brain which is highly expressed. There is a close association between susceptibility of infection to JCV and expression of this DNA binding protein. We have made expression vectors for all four class members of the NF-1 family. We have made a unique human brain derived, multipotential progentior cell line that can differentiate to either neurons or glial cells. We have been able to over express NF-1/X in human neurons differentiated from human CNS progenitor cells that are not susceptible to infection since these cells do not express competent levels of NF-1/X. The NF-1/X over expressing neurons however are susceptible to JCV infection, confirming the necessary regulatory function of this transcription factor as we have shown in the past in hematopoietic cells. In addition, we have shown that the binding site for NF-1/X is directly adjacent to the c-jun binding site on the viral promoter. We have then shown that NF-1/X and c-jun proteins interact directly. Increased concentrations of c-jun can alter the ability of NF-1/X to bind to its cognate site particularly if c-jun is phosphorylated. We have also determined that JCV infection of human multiotential progenitor cells results in viral multiplication that is augmented 10 fold when differentiated to astrocytes but shut off when differentiated to neurons. New experiments have shown that infection can be reversed if the lineage is directed either back to progenitor cells or from glial cells to neurons. There is also an interplay among the class members of NF-1 since if increases of NF-1A in cultured cells diminishes JCV growth while its unique inhibition compared with NF-1X shows an increase in JCV growth. As a part of these studies, we have demonstrated that virus infection kills human glial cells as a lytic, necrotic cell death and not an apoptotic cell death as previously described by others. We have also been able to direct differentiation of the progenitor cells to an oligodendrocyte. However, these cells are not as susceptible to JCV productive infection as the astrocytes due in some measure to their lack of high levels of expression of DNA binding proteins that recognize the viral promoter. Also few of the oligodendroctes express the viral co-receptor, 5HT2A. We have also identified the biological effects of TGF-beta that appears to augment the ability of JCV to multiply. TGF-beta also appears to increase the positive effects of the transcriptionally active proteins, Smads. Inhibitors of TGF beta or activity of Smads block the augmentation of JCV growth. This may also be true for the other human polyomavirus BKV, the cause of kidney allograft failure in about 3% of recipients. Interestingly, in another study, we found that high antiviral antibody titers to BKV in kidney donors correlates with graft failure in the recipient. It may be important to investigate the role of TGF beta in this disease process as well. In addition to these factors, we have identified another DNA binding protein, SpiB, that is upregulated in patients on therapies that are a risk for PML i.e. natalizumab. The SpiB protein functions in lymphoid cells as a regulator for B cell development and also binds sites in the JCV promoter. These sites are overexpressed in the nucleotide sequences that are found in the virus DNA in PML brain tissue while not found in the same number in non pathologic tissue like kidney.
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