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. 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. As a CLIA certified laboratory we have also participated in collaborative, viral diagnostic work that identified the human BK polyomavirus assoicated with allograft recipients published in the New England Journal of Medicine. We also have surveyed over 1500 serum samples and determined that kidney donors with high antibody titers to BKV may pass on the infection to the recipient. We have also conducted a large scale review of patient CSF and plasma samples from over 400 MS patients and normal aged and gender matched controls to determine whether JCV DNA is present in PML patients. This has been part of an evaluation of the risk of MS patients treated with natalizumab, a humanized monoclonal antibody that prevents T cell migration into the brain. It is clear that JCV DNA can be found only in the CSF of PML patients further validating the PCR analysis of CSF as a diagnostic tool for PML. There is evidence however of a persistent viremia in a small number of individuals who maintain presence of JCV DNA in their peripheral circulation for months.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Intramural Research (Z01)
Project #
1Z01NS001983-35
Application #
7322924
Study Section
(LMMN)
Project Start
Project End
Budget Start
Budget End
Support Year
35
Fiscal Year
2006
Total Cost
Indirect Cost
City
State
Country
United States
Zip Code
Major, Eugene O (2009) Progressive Multifocal Leukoencephalopathy in Patients on Immunomodulatory Therapies. Annu Rev Med :
Linda, Hans; von Heijne, Anders; Major, Eugene O et al. (2009) Progressive multifocal leukoencephalopathy after natalizumab monotherapy. N Engl J Med 361:1081-7
Major, Eugene O (2009) Reemergence of PML in natalizumab-treated patients--new cases, same concerns. N Engl J Med 361:1041-3
Bohl, Daniel L; Brennan, Daniel C; Ryschkewitsch, Caroline et al. (2008) BK virus antibody titers and intensity of infections after renal transplantation. J Clin Virol 43:184-9
Houff, Sidney A; Berger, Joseph; Major, Eugene O (2008) Response to Linberg et al. Natalizumab alters transcriptional expression profiles of blood cell subpopulations of multiple sclerosis patients. J Neuroimmunol 199:160-161
Roseti, Livia; Facchini, Andrea; De Franceschi, Luciana et al. (2007) Induction of original phenotype of human immortalized chondrocytes: a quantitative gene expression analysis. Int J Mol Med 19:89-96
Ravichandran, Veerasamy; Jensen, Peter N; Major, Eugene O (2007) MEK1/2 inhibitors block basal and transforming growth factor 1beta1-stimulated JC virus multiplication. J Virol 81:6412-8
Ravichandran, Veerasamy; Major, Eugene O (2006) Viral proteomics: a promising approach for understanding JC virus tropism. Proteomics 6:5628-36
Li, Jongming; Melenhorst, Jos; Hensel, Nancy et al. (2006) T-cell responses to peptide fragments of the BK virus T antigen: implications for cross-reactivity of immune response to JC virus. J Gen Virol 87:2951-60
Yousry, Tarek A; Major, Eugene O; Ryschkewitsch, Caroline et al. (2006) Evaluation of patients treated with natalizumab for progressive multifocal leukoencephalopathy. N Engl J Med 354:924-33

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