Detection of archetype and rearranged variants of JC virus in multiple tissues from a pediatric PML patient

Rearrangements of the JC virus (JCV) regulatory region (RR) are consistently found in the brains of patients with progressive multifocal leukoencephalopathy (PML), whereas the archetype RR is present in their kidneys. In addition, the C terminus of the large T antigen (T-Ag) shows greater variability in PML than does the rest of the coding region. To determine whether similar changes in simian virus 40 (SV40) are necessary for disease induction in monkeys, we sequenced the SV40 RR and the C terminus of the T-Ag from the brain of simian/human immunodeficiency virus (SHIV)-infected monkey 18429, which presented spontaneously with an SV40-associated PML-like disease, as well as from the peripheral blood mononuclear cells (PBMC), kidneys, and brains of SV40-seronegative, SHIV-infected monkeys 21289 and 21306, which were inoculated with the 18429 brain SV40 isolate. These animals developed both SV40-associated PML and meningoencephalitis. Thirteen types of SV40 RR were characterized. Compared to the SV40 archetype, we identified RRs with variable deletions in either the origin of replication, the 21-bp repeat elements, or the late promoter, as well as deletions or duplications of the 72-bp enhancer. The archetype was the most prominent RR in the brain of monkey 18429. Shortly after inoculation, a wide range of RRs could be found in the PBMC of monkeys 21289 and 21306. However, the archetype RR became the predominant type in their blood, kidneys, and brains at the time of sacrifice. On the contrary, the T-Ag C termini remained identical in all compartments of the three animals. These results indicate that unlike JCV in humans, rearrangements of SV40 RR are not required for brain disease induction in immunosuppressed monkeys.Progressive multifocal leukoencephalopathy (PML) is emerging as a persistent cause of death in human immunodeficiency virus-infected patients, despite the availability of highly active antiretroviral therapy, and survival is limited to an average of 11 months (1). This disease is the consequence of a lytic infection of oligodendrocytes by the polyomavirus JC (JCV). Since there is no specific treatment for PML, the development of an animal model of PML would be a tremendous asset for studying its pathophysiology and devising therapeutic interventions. However, earlier attempts to create a useful animal model of PML were unsuccessful since the host range of JCV is restricted to humans. Indeed, this virus may cause a variety of tumors in animals but it does not replicate in glial cells of rodents or nonhuman primates (9, 19, 23), probably because of a block at the transcriptional level (3).Since JCV cannot be used directly to create an animal model of PML in immunosuppressed monkeys, we hypothesized that a closely related simian polyomavirus could be used to this effect. Simian polyomavirus 40 (SV40) has 69% sequence homology with JCV, and a few cases of PML-like disease have been reported in rhesus monkeys infected with SV40, with detection of polyomavirus-like virions or SV40 nucleic acid and large ...

Section: Discussionmentioning

confidence: 92%

Rearrangement of Simian Virus 40 Regulatory Region Is Not Required for Induction of Progressive Multifocal Leukoencephalopathy in Immunosuppressed Rhesus Monkeys

Dang

Axthelm

Letvin

et al. 2005

“…While tandem repeat variants have been found in the central nervous system of patients with PML, in brain tumors, and in peripheral lymphocytes of both PML and non-PML patients, the archetype has rarely been found in diseased brains. These findings suggest that the presence of a specific strain might reflect viral adaptation to the particular host tissue (4,16,19,22).Because the Mad-1 strain has the capacity for lytic infection of oligodendrocytes, JCV has always been considered strictly neurotropic. However, recent studies have shown that certain strains of JCV can replicate in lymphoid cells and that the capability of the virus to replicate in host cells is highly dependent on the structure of the TCR (2, 14).…”

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confidence: 88%

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confidence: 99%

Mad-1 Is the Exclusive JC Virus Strain Present in the Human Colon, and Its Transcriptional Control Region Has a Deleted 98-Base-Pair Sequence in Colon Cancer Tissues

Ricciardiello

Chang

Laghi

et al. 2001

JC virus (JCV), along with other members of the polyomavirus family, encodes a class of highly conserved proteins, T antigens, that are capable of inducing aneuploidy in cultured cells. We have previously isolated T-antigen DNA variants of JCV from both colon cancer tissues and the corresponding nonneoplastic gastrointestinal tissues, raising new questions about the role of JCV in the development of chromosomal instability of the colon. Based on the sequence of the transcriptional control region (TCR), JCV can be classified as archetype or tandem repeat variants. Among the latter, Mad-1, the prototype virus first isolated from a patient with progressive multifocal leukoencephalopathy, is characterized by lacking the 23-and 66-bp sequences that are present in the archetype and by duplication of a 98-bp sequence. In this study, we evaluated differences in the TCR of JCV isolated from colon cancer tissues and nonneoplastic epithelium. To characterize JCV variants, we first treated eight pairs of DNA samples from colon cancers and noncancerous tissue with topoisomerase I and then amplified and cloned the JCV TCR. We obtained 285 recombinant clones from the JCV TCR, 157 from nonneoplastic samples, and 128 from colon cancer tissues. Of these clones, 262 spanned the length of the JCV Mad-1 TCR: 99.3% from nonneoplastic samples and 82.8% from colon cancer tissues. In sequencing 54 clones in both directions, we did not find archetype JCV either in the nonneoplastic tissue or in the cancer samples. From all colon cancer tissues, 18 clones had a deletion of one 98-bp tandem repeat. This deleted strain was not detected in any of the nonneoplastic tissues (14 versus 0% [ 2 ‫؍‬ 23.6; P < 0.001]). Our study demonstrates that the only JCV strain present in the human colon is Mad-1, and the variant with a single 98-bp sequence is found exclusively in the cancer tissues. This strain may be involved in the development of chromosomal instability.The human polyomavirus JC virus (JCV) infects a large proportion of the population worldwide (23). The initial infection is unapparent, but subsequently JCV establishes a latent infection, becoming reactivated when the immune system is impaired (8). The JCV genome is double-stranded, negatively supercoiled circular DNA that consists of early and late coding regions separated by two noncoding regions at both the 5Ј and 3Ј ends. Within the noncoding sequences is the transcriptional control region (TCR), which contains the promoter and enhancer for the early and late proteins; the intergenic region separates the 3Ј regions of T antigen and the capsid protein, VP1.T antigen is an oncogenic protein capable of transforming mammalian cells by interacting with cellular proteins such as p53 and the Rb family proteins (5, 21). T antigen also has ATPase and helicase activities, the latter of which may eventually contribute to chromosomal breakage and recombination (7). We have recently reported the presence of JCV T-antigen DNA sequences in nonneoplastic gastrointestinal tissues, colon cancer samples,...

“…Although the etiologic role of JCV in PML is well documented, the pathogenesis and, in particular, the role of viral determinants is less clear. Virtually all JCV strains isolated from the brain or CSF of PML patients are characterized by highly variable genomic rearrangements of the non-coding control region (NCCR), which governs viral early and late genes in opposite directions of the circular polyomavirus DNA genome (1,4,31,39,41,43,49,54,59). In contrast, JCV detected in the urine of immunocompetent individuals show a consistent linear architecture called the archetype NCCR (at-NCCR).…”

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confidence: 99%

Rearranged JC Virus Noncoding Control Regions Found in Progressive Multifocal Leukoencephalopathy Patient Samples Increase Virus Early Gene Expression and Replication Rate

Gosert

Kardas

Major

et al. 2010

117

145

Polyomavirus JC (JCV) infects ϳ60% of the general population, followed by asymptomatic urinary shedding in ϳ20%. In patients with pronounced immunodeficiency, including HIV/AIDS, JCV can cause progressive multifocal leukoencephalopathy (PML), a devastating brain disease of high mortality. While JCV in the urine of healthy people has a linear noncoding control region called the archetype NCCR (at-NCCR), JCV in brain and cerebrospinal fluid (CSF) of PML patients bear rearranged NCCRs (rr-NCCRs). Although JCV NCCR rearrangements are deemed pathognomonic for PML, their role as a viral determinant is unclear. We sequenced JCV NCCRs found in CSF of eight HIV/AIDS patients newly diagnosed with PML and analyzed their effect on early and late gene expression using a bidirectional reporter vector recapitulating the circular polyomavirus early and late gene organization. The rr-NCCR sequences were highly diverse, but all increased viral early reporter gene expression in progenitor-derived astrocytes, glia-derived cells, and human kidney compared to the expression levels with the at-NCCR. The expression of simian virus 40 (SV40) large T antigen or HIV Tat expression in trans was associated with a strong increase of at-NCCR-controlled early gene expression, while rr-NCCRs were less responsive. The insertion of rr-NCCRs into the JCV genome backbone revealed higher viral replication rates for rr-NCCR compared to those of the at-NCCR JCV in human progenitor-derived astrocytes or glia cells, which was abrogated in SV40 large T-expressing COS-7 cells. We conclude that naturally occurring JCV rr-NCCR variants from PML patients confer increased early gene expression and higher replication rates compared to those of at-NCCR JCV and thereby increase cytopathology.