Human Polyomavirus JC monitoring and noncoding control region analysis in dynamic cohorts of individuals affected by immune-mediated diseases under treatment with biologics: an observational study

Abstract

BackgroundProgressive multifocal leukoencephalopathy (PML) onset, caused by Polyomavirus JC (JCPyV) in patients affected by immune-mediated diseases during biological treatment, raised concerns about the safety profile of these agents. Therefore, the aims of this study were the JCPyV reactivation monitoring and the noncoding control region (NCCR) and viral protein 1 (VP1) analysis in patients affected by different immune-mediated diseases and treated with biologics.MethodsWe performed JCPyV-specific quantitative PCR of biological samples collected at moment of recruitment (t0) and every 4 months (t1, t2, t3, t4). Subsequently, rearrangements’ analysis of NCCR and VP1 was carried out. Data were analyzed using χ2 test.ResultsResults showed that at t0 patients with chronic inflammatory rheumatic diseases presented a JCPyV load in the urine significantly higher (p≤0.05) than in patients with multiple sclerosis (MS) and Crohn’s disease (CD). It can also be observed a significant association between JC viruria and JCPyV antibodies after 1 year of natalizumab (p=0.04) in MS patients. Finally, NCCR analysis showed the presence of an archetype-like sequence in all urine samples, whereas a rearranged NCCR Type IR was found in colon-rectal biopsies collected from 2 CD patients after 16 months of infliximab. Furthermore, sequences isolated from peripheral blood mononuclear cells (PBMCs) of 2 MS patients with JCPyV antibody at t0 and t3, showed a NCCR Type IIR with a duplication of a 98 bp unit and a 66 bp insert, resulting in a boxB deletion and 37 T to G transversion into the Spi-B binding site. In all patients, a prevalence of genotypes 1A and 1B, the predominant JCPyV genotypes in Europe, was observed.ConclusionsIt has been important to understand whether the specific inflammatory scenario in different immune-mediated diseases could affect JCPyV reactivation from latency, in particular from kidneys. Moreover, for a more accurate PML risk stratification, testing JC viruria seems to be useful to identify patients who harbor JCPyV but with an undetectable JCPyV-specific humoral immune response. In these patients, it may also be important to study the JCPyV NCCR rearrangement: in particular, Spi-B expression in PBMCs could play a crucial role in JCPyV replication and NCCR rearrangement.

JCPyV NCCR analysis in PML patients with different risk factors: exploring common rearrangements as essential changes for neuropathogenesis

“…The resulting supernatants were used directly in molecular biology assays. To detect and quantify the JCPyV genome copy numbers, a 7300 Real-Time quantitative PCR (qPCR) system (Applied Biosystems, USA) was used employing specific primers and probes [18]. Nested PCR with specific primers flanking the NCCR and VP1 regions were performed and PCR products corresponding to JCPyV NCCR and VP1 regions were purified with the QIAquick PCR purification kit, according to QIAGEN protocol [19].…”

Lymphocyte Gene Expression and JC Virus Noncoding Control Region Sequences Are Linked with the Risk of Progressive Multifocal Leukoencephalopathy

“…Spi-B levels are higher in cells permissive to JC virus (JCV) transcription and replication, and overexpression of Spi-B increases viral gene expression (8). Spi-B binds to target sites in the JCV noncoding control region (NCCR) isolated from PML brain tissue but not in the archetype NCCR commonly detected in the urine of asymptomatic healthy individuals (8)(9)(10)(11). Importantly, mutation of these sites in PML-associated NCCRs decreases Spi-B protein binding and viral gene expression (8,12).…”

Natalizumab-treated patients at high risk for PML persistently excrete JC polyomavirus

“…1) (www.biogen.com), augmenting risk algorithms with additional risk factors seems imperative. The prevalence of JCV viruria among natalizumab-treated patients is reported to cover a wide range (18–87 %) (Delbue et al 2015; Lanzillo et al 2014; Laroni et al 2012; Bellizzi et al 2013; Rinaldi et al 2010), but no association between viruria prevalence and progression to PML has been found. The strong correlation demonstrated herein between AI and JCV viruria indicates that natalizumab-treated patients with active JCV infection commonly fall into a high-risk category for progression to clinical disease (Plavina et al 2014).…”

“…Intermittent excretion made the measure of urinary JCV of little diagnostic benefit, leaving it largely ignored in the approach to mitigating PML risk. Given that appropriate interrogation of the urine reveals a consistent signal that likely represents encapsidated virus, that the viral load is high and strongly correlated with high antibody titer, and the fact that the level of viruria increases with progression to PML (Delbue et al 2015; Bellizzi et al 2013; Domínguez-Mozo et al 2015) suggest urinary JCV has clinical value. Active JCV infection in the urine can often be detected prior to JCV antibodies (Laroni et al 2012; Lanzillo et al 2014), directing attention to those patients that may already be heading down the path to neurotropic JCV.…”