Role Of JC Virus Research Articles

Presence of JC Virus DNA in the Tumor Tissue and Normal Mucosa of Patients with Sporadic Colorectal Cancer (CRC) or with Positive Family History and Bethesda Criteria

JC virus (JCV) may infect the gastrointestinal tract in childhood, and, by encoding a gene for T-antigen (T Ag), can initiate chromosomal instability in epithelial cells. We looked for JCV DNA in the cancer tissue of patients with sporadic colorectal cancer (CRC, Group A) and with positive family history and Bethesda criteria (Group B). We hypothesized that the role of JCV may be different between these two groups. Fifty-six patients were randomly selected from our database, 30 in Group A and 26 in Group B. DNA was isolated from the tumor, normal mucosa, and plasma, and JCV DNA sequences were looked for with specific polymerase chain reaction (PCR) assays for T Ag primers. Immunohistochemistry for hMLH1, hMSH2, hMSH6, and PMS2 was performed on paraffin-embedded tissue. In Group A, T Ag was demonstrated in 6 (20.00%) and 3 (10.00%) of the tumors and adjacent normal mucosa, respectively (P = 0.094). In Group B, the corresponding observations were 10 (38.46%) and 6 (23.07%), respectively (P < 0.001). Immunohistochemistry for hMLH1, hMSH2, hMSH6, and PMS2 was performed in all of the Group A and B patients. All patients of Group A (100%) showed expression of these proteins, while only 19 patients of Group B did so (73.1%), P = 0.009. JCV T Ag DNA was found in 20, 28.5, and 42.1% of the tumors in Group A, Group B with negative staining for DNA repair genes, and Group B with a positive staining, respectively (NS). CRC patients with positive family history have a higher incidence of JCV T Ag, but this did not correlate with specific DNA repair gene mutations. We could not conclude that, on the background of genetic mutation in one of the DNA repair genes, JCV acts as the missing link in the chain of events leading to CRC.

Announcing the winner of the 2010 Kurt Jellinger Prize

Dr. Yukiko Shishido-Hara is the winner of the 2010 Kurt Jellinger Prize for Excellence in Scientific Writing, based on the review article ‘‘Progressive multifocal leukoencephalopathy and promyelocytic leukemia nuclear bodies: a review of clinical, neuropathological, and virological aspects of JC virus-induced demyelinating disease’’, published in this issue of Acta Neuropathologica and reflecting the author’s experience in both neuropathology and virology. Yukiko Shishido-Hara has been engaged in the study of JC virus since early research as a fellow at the National Institutes of Health, Tokyo and the Tokyo Metropolitan Institute for Neuroscience (1992–1995), where she examined the expression of JC virus proteins in insect cells using Baculoviral vectors and in COS7 cells using the SRa promoter. From 1995 to 1999, she continued her studies at the NIH, USA, by examining JC virus in AIDS patients, and by determining the complete sequence of JC virus Tokyo-1 strain. After moving back to Tokyo, she has explored a variety of projects related to JC virus, including the induction of oligodendroglial cells from cultured neural stem cells, the nuclear structure of JC virus-infected human oligodendroglial cells, the role of JC virus in brain tumors, and the pathogenic role of promyelocytic leukemia nuclear bodies (PML-NBs) as an intranuclear target of JC virus infection, which represents a major part of her Prize winning paper. Yukiko Shishido-Hara earned MD and PhD degrees, and she underwent training in both virology and neuropathology before obtaining certification in pathology. Currently, she serves as surgical pathologist and Assistant Professor, Department of Pathology, Kyorin University School of Medicine, Tokyo and as Visiting Scientist, Tokyo Metropolitan Institute for Neuroscience. The 2010 Kurt Jellinger Prize includes the invitation to the 17th International Congress of Neuropathology in September 2010 (ICN 2010) organized by Herbert Budka and Hans Lassmann from Vienna, a capital city of neuropathology where Acta Neuropathologica has been founded almost 50 years ago and edited for 43 years by Franz Seitelberger und Kurt Jellinger. The Prize also includes the invitation to join the Editorial Board which represents a group of enthusiastic scientists wishing to provide the best service to the neurocommunity. The present issue includes review papers written by speakers of ICN 2010. To make these and other papers as topical as possible, processing of papers is very fast in this journal. For example, the mean time from submission to first decision has been 12 days in 2010, and it was 3 days for the ICN 2010 papers included in this issue. This has been possible only thanks to a dynamic Editorial Board which is glad to have Yukiko Shishido-Hara and her expertise aboard. W. Paulus (&) Institute of Neuropathology, University Hospital Munster, Domagkstr. 19, 48149 Munster, Germany e-mail: werner.paulus@uni-muenster.de

The presence of JC virus in gastric carcinomas correlates with patient's age, intestinal histological type and aberrant methylation of tumor suppressor genes

JC virus (JCV) is a neurotropic polyomavirus and the causative agent of progressive multifocal leukoencephalopathy. A role for JCV in gastrointestinal malignancies has been recently suggested. This study was carried out to determine the prevalence of polyomaviruses including JCV, BKV and SV40 in gastric cancers in Tunisia and to determine the clinicopathological characteristics of virus-associated gastric carcinomas. The presence of polyomaviruses DNA sequences was surveyed in 61 cases of primary gastric carcinomas and in 53 paired non-tumor gastric mucosa by PCR. Findings were correlated to clinicopathological parameters, p53 expression and methylation status of 11 tumor-related genes. Using PCR assays, JCV T-antigen sequence was more frequently detected in gastric carcinomas than in non-tumor gastric mucosa (26 vs 6%, P=0.03), while those of SV40 and BKV were not detected in any cases. Correlation analysis showed that JCV had higher frequency in patients older than 55 years (P=0.034) and in the intestinal histological type (P=0.04). With regard to methylation status, P16 and P14 showed significantly higher methylation frequencies in JCV-positive gastric carcinomas than in JCV-negative cases (P=0.007 and P=0.003, respectively). Moreover, the mean of the methylation index was significantly higher in JCV-positive than in JCV-negative cases (P=0.024). In multivariate logistic regression analysis, age of patients and the methylation index are only the two independent factors associated with JCV infection. Kaplan–Meier survival analysis showed a trend toward better survival for JCV-associated gastric carcinomas patients (log-rank, P=0.11). Our study suggests a role of JCV as cofactor in the pathogenesis of the intestinal type of gastric carcinomas in older persons.

Viral infection in bone marrow transplants: Is JC virus involved?

Hemorrhagic cystitis is characterized by hematuria due to inflammation of the bladder. In bone marrow transplants, this disease is linked to the infection by human polyomavirus BK, whereas the role of the human polyomavirus JC is unclear. The transcriptional control regions of both viruses contain important cellular transcription factor binding sites that undergo rearrangement process generating suitable variants that could be more active for viral replication and for the onset of hemorrhagic cystitis. In this study urine obtained from seven patients with bone marrow transplant were examined. Polyomavirus genomes were quantified by PCR and viral loads were compared. The transcriptional regions of both viruses were amplified and sequenced to determine the presence of variants. Subtypes of polyomaviruses were determined by amplification and sequencing of the viral protein 1 region. The results showed that four of seven patients were positive for BK DNA, two of seven patients had BK and JC DNA and one of seven had JC DNA. Positive samples were amplified and sequenced successively for transcriptional regions. The viral archetype was always found in both viruses. Finally, typing showed that BK virus subtype I infected patients with BK, whereas JC virus genotype IA and genotype 1B were found in patients infected with JC. The data suggest that new and different approaches are required to improve the morbidity and mortality caused by polyoma-associated hemorrhagic cystitis, since it known that BK virus is involved in the onset of hemorrhagic cystitis, whereas the role of JC virus should be investigated further.

Polyomavirus BK and JC infections in well matched Finnish kidney transplant recipients

Since 2003, only one case of polyomavirus-associated nephropathy (PVAN) has occurred in our clinic despite screening protocols. In contrast to BK virus, the role of JC virus in PVAN is unclear. We studied the incidence and impact of polyomavirus BK and JC viruria and PVAN in well-matched Finnish kidney transplant recipients. All Helsinki University Hospital kidney transplant recipients between 2004 and 2006 were prospectively followed (n = 163). Patients with a 12-month protocol-biopsy taken and polyomavirus urinary secretion screened by PCR were studied (n = 68). Cyclosporine-based triple-drug immunosuppression was usually used. BK or JC viruria was detected in 18 (27%) and 14 (21%) patients after transplantation respectively. Persistent BK or JC viruria was found in 5 (7%) and 9 (13%) patients. No cases of PVAN were diagnosed from protocol biopsies or from biopsies taken for clinical indications. A positive BK or JC viruria or persistent viruria was not associated with reduced renal function at follow-up, histopathologic changes in 12-month protocol biopsies, or acute rejections. The incidence of BK and JC viruria was similar to what has been previously reported, but no cases of polyomavirus-associated nephropathy were seen in our well-matched kidney transplant population.

S1993 The Role of JC Virus in Colorectal Neoplasia Occurring After Liver Transplantation

S1993 The Role of JC Virus in Colorectal Neoplasia Occurring After Liver Transplantation

Longitudinal study of two cases of progressive multifocal leukoencephalopathy with a clinical benign evolution

Progressive multifocal leukoencephalopathy (PML) usually is a rapid and fatal demyelinating disease of the central nervous system (CNS), caused by JC virus (JCV). After the introduction of Highly active antiretroviral therapy (HAART), its prognosis has been modified in some cases but remains a relevant cause of morbidity in human immunodeficiency virus-seropositive (HIV+) patients. The authors report here two cases of PML, followed over time, sharing a benign course and a JCV antigen-specific T-cell response, but with different cerebrospinal fluid (CSF), magnetic resonance imaging (MRI), and clinical features. In both cases, JCV DNA detection in brain biopsies samples and specific antigenic response preceded its isolation in the CSF by several months. In one patient, during the first stage of the disease, the presence of CSF and MRI inflammatory findings, associated with the lack of JCV detection in the CSF, made the diagnosis more challenging. Given that to date a reformation of the laboratory parameters for PML diagnosis is strongly needed, this report highlights the following considerations: (a) indications for performing brain biopsy in HIV-related leukoencephalopathies of uncertain origin, and (b) the role of JCV immunologically specific T-cell response as an additional marker for PML diagnosis and indicator for good prognosis of the disease.

Rarity of JC virus DNA sequences and early proteins in human gliomas and medulloblastomas: the controversial role of JC virus in human neurooncogenesis

JC virus (JCV), the agent of progressive multifocal leucoencephalopathy (PML), exerts an oncogenic effect in several laboratory animal models. Moreover, JCV genomic DNA and early viral protein T-antigen have been detected in various types of human central nervous system (CNS) neoplasms. To further explore this association we have studied paraffin-embedded brain biopsy tissue from 60 neoplasms (55 gliomas and five medulloblastomas) and 15 reactive gliosis cases for the presence of JCV DNA sequences and proteins. Four post mortem cases of HIV-associated PML were used as positive controls. Samples were assessed by polymerase chain reaction (PCR) amplification of early (large T antigen) and late (virion protein 3) sequences and immunohistochemistry (IHC) with both PAb 2024 and anti-SV40 large T antigen monoclonal antibodies. Five cases (three neoplasms and two reactive gliosis instances) showed low viral DNA levels when PCR-tested for VP3 or large T, while no case was immunoreactive for any of the two antibodies used. The four PML cases yielded positive results with both PCR and IHC. Additionally, IHC with both antibodies was applied to a tissue micro-array including 109 CNS tumours and 21 reactive gliosis samples. No immunoreactivity was detected in any of these tissue micro-array samples. The rarity of JCV DNA sequences and early proteins in our brain tumours enriches the controversy over the role of JCV in human neurooncogenesis, whose clarification is in need of further molecular and epidemiologic studies.

JC virus small tumor antigen promotes S phase entry

A number of studies revealed the detection of human polyomavirus, JC virus (JCV), in a variety of human tumors. Viral large tumor antigen (T-Ag) is known to play a major role in cell proliferation and transformation but the role of JCV small t-Ag in such processes remains completely unknown. Several studies from monkey polyomavirus, SV40, small tumor antigen (tAg) suggest that it significantly contributes to the large T-Ag-mediated cell transformation. Here, we investigated the effects of JCV small t-Ag on cell cycle progression and cell proliferation. Examination of subcellular distribution of small t-Ag by means of immunocytochemistry demonstrated that it mostly localizes to the cytoplasmic compartment of the cell. Cell cycle progression analysis of a human glioblastoma cell line, U-87MG, demonstrated that small t-Ag expressing cells enter S phase of the cell cycle at a significantly higher rate than controls when released from G0/G1 arrest. In addition, the growth rate of these cells is consistent with the rate of cell cycle progression and is significantly enhanced compared to controls. Moreover, analysis of the mechanism through which small t-Ag accelerates the cell cycle progression showed that JCV small t-Ag targets a major serine/threonine phosphatase, PP2A, and its interaction domain with this enzyme is mapped toward the middle portion of the protein, residues 82 to 124. Furthermore, examination of the cell cycle stage specific expression profiles of selected cyclins and cyclin-dependent kinases, particularly those active in G1/S and G2/M transition states demonstrated that the rate of the appearance of cyclin E, cyclin B, and cyclin E-dependent kinase 2 (Cdk2) substantially increased in small t-Ag positive cells as compared to controls when the cells were released from G0/G1 arrest. In parallel, in vitro analysis of cyclin E-associated H1 kinase activity of clones demonstrated that small t-Ag positive clones exhibited significantly higher activity than controls. Additionally, results from reporter gene assays showed that small tAg has the ability to activate the cyclin E promoter which is consistent

Molecular characterisation of JC virus strains detected in human brain tumours

The aim of this study was to evaluate the presence and significance of JC virus (JCV) in human brain tumours. Histology, immunohistochemistry (IHC) and molecular biology techniques were employed to examine specimens of tumour tissue, peripheral blood and cerebrospinal fluid taken from 22 patients with primary neuro-epithelial tumours. Furthermore, the coding viral protein (VP1) region and non-coding transcription control region (TCR) of JCV genome isolated from the tumours were submitted to sequence analysis in order to detect viral rearrangements or mutations. JCV genome was found in nine of the 22 tumour specimens (40.9%), including eight astrocyte-derived tumours (seven glioblastomas and one astrocytoma) and one oligodendroglioma, and in two of the 15 cerebrospinal fluid specimens (13.3%) with positive tumour tissue (one glioblastoma and one astrocytoma). Sequence analysis of JCV VP1, which was amplified in seven tissue samples and the two cerebrospinal fluid samples, revealed only genotype 1 (four 1a and three 1b), whereas TCR was amplified in six tissue samples and only one cerebrospinal fluid sample. TCR sequence analysis was possible in four cases and identified three Mad-4 and one type II sequences; the TCR genomic structures of JCV isolated from cerebrospinal fluid were the same as those sequenced from corresponding tumour tissue, thus indicating a possible cerebrospinal fluid dissemination of neoplastic cells carrying viral DNA. Our results suggest a possible role of JCV in the induction of brain tumours, especially in those originating from brain cells normally targeted by JCV infection.

Association of prolonged survival in HLA-A2+ progressive multifocal leukoencephalopathy patients with a CTL response specific for a commonly recognized JC virus epitope.

The role of JC virus (JCV)-specific CTL was explored in the immunopathogenesis of progressive multifocal leukoencephalopathy (PML). We identified a 9-aa epitope of the JCV capsid protein VP1, the VP1(p100) peptide ILMWEAVTL, which is recognized by CTL of HLA-A2+ HIV+/PML survivors. We then constructed an HLA-A*0201/VP1(p100) tetrameric complex that allowed us to assess by flow cytometry the PBMC of 13 PML patients and 11 control subjects for the presence of JCV-specific CTL. VP1(p100)-specific CTL were detected by tetramer binding in VP1(p100)-stimulated PBMC of five of seven (71%) PML survivors and zero of six PML progressors (p = 0.02). Two of three HIV+ patients with a leukoencephalopathy resembling PML, but with no virologic evidence of JCV infection, also had detectable VP1(p100)-specific CTL in their PBMC. PBMC of eight HIV+ patients with other neurologic diseases and healthy control subjects had no detectable JCV-specific CTL. These data suggest that the JCV-specific cellular immune response may be important in the containment of PML, and the tetramer-staining assay may provide a useful prognostic tool in the clinical management of these patients.

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.

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% [chi(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.

Human polyomavirus JC control region variants in persistently infected CNS and kidney tissue.

The question of a possible role for JC virus (JCV) genomic rearrangements in the pathogenesis of progressive multifocal leukoencephalopathy (PML) was addressed by analysis of the genomic complexity and the transcriptional control region (TCR) of the JCV DNA population in persistently infected CNS and kidney tissue. After cloning of full-length viral DNA, no extensive changes were detected in the coding regions of the JCV genome by restriction analysis suggesting an intact JCV DNA population. For further analysis of the distribution of JCV subtypes, the non-coding region was amplified by PCR. Molecular analysis revealed homogeneous JCV TCR populations in almost 50% of the individuals. Heterogeneity was found in two CNS samples with three and five different JCV subtypes, respectively, and in four kidney specimens with two TCR subtypes. Altogether, seven TCR subtypes were identified. One in each group represented single promoter element TCRs without duplication of sequences. The TCR of the major variant JCV-W1 was comparable in sequence and structure to that of the PML prototype JCV Mad-1 DNA. The identification of dominant PML-derived JCV TCR subtypes in most persistently infected individuals suggests that rearrangements of the JCV TCR can be associated with the persistent state of infection. However, it appears unlikely that PML-associated JCV subtypes are generated anew in each individual host in the course of persistence. The findings rather suggest that a limited number of stable JCV subtypes circulate in different geographical regions of the world.

Tumor pathogenesis of human neurotropic JC virus in the CNS.

In the last several years, studies have reported the detection of sequences similar to the polyomavirus, Simian Virus 40 (SV40) in human tumors including choroid plexus papillomas and ependymomas. Taken together with well-established evidence that SV40, as well as the human polyomaviruses JC virus (JCV) and BK virus (BKV), are oncogenic in several animal models, new interest has resurfaced regarding a possible role for these viruses in human tumors. In particular, a strong case can be made for re-evaluating the oncogenic potential of JCV in a neurotropic polyomavirus which is the causative agent of the fatal human demyelinating disease, Progressive Multifocal Leukoencephalopathy (PML). In this review, we discuss the transforming capability of JCV in vitro, JCV's ability to induce neural origin tumors in non-human primates as well as in rodents, and several case reports which suggest a possible role for JCV in tumor pathogenesis in the central nervous system.