Luiz Henrique da Silva Nali

High prevalence of the simultaneous excretion of polyomaviruses JC and BK in the urine of HIV-infected patients without neurological symptoms in São Paulo, Brazil

OBJECTIVE To evaluate the prevalence of the urinary excretion of BKV and JCV in HIV-infected patients without neurological symptoms. METHODS Urine samples from HIV-infected patients without neurological symptoms were tested for JC virus and BK virus by PCR. Samples were screened for the presence of polyomavirus with sets of primers complementary to the early region of JCV and BKV genome (AgT). The presence of JC virus or BK virus were confirmed by two other PCR assays using sets of primers complementary to the VP1 gene of each virus. Analysis of the data was performed by the Kruskal-Wallis test for numerical data and Pearson or Yates for categorical variables. RESULTS A total of 75 patients were included in the study. The overall prevalence of polyomavirus DNA urinary shedding was 67/75 (89.3%). Only BKV DNA was detected in 14/75 (18.7%) urine samples, and only JCV DNA was detected in 11/75 (14.7%) samples. Both BKV and JCV DNA were present in 42/75 (56.0%) samples. CONCLUSION In this study we found high rates of excretion of JCV, BKV, and simultaneous excretion in HIV+ patients. Also these results differ from the others available on the literature.

Genomic analysis of ERVWE2 locus in patients with multiple sclerosis: absence of genetic association but potential role of human endogenous retrovirus type W elements in molecular mimicry with myelin antigen

Human endogenous retroviruses (HERVs) arise from ancient infections of the host germline cells by exogenous retroviruses, constituting 8% of the human genome. Elevated level of envelope transcripts from HERVs-W has been detected in CSF, plasma and brain tissues from patients with Multiple Sclerosis (MS), most of them from Xq22.3, 15q21.3, and 6q21 chromosomes. However, since the locus Xq22.3 (ERVWE2) lack the 5′ LTR promoter and the putative protein should be truncated due to a stop codon, we investigated the ERVWE2 genomic loci from 84 individuals, including MS patients with active HERV-W expression detected in PBMC. In addition, an automated search for promoter sequences in 20 kb nearby region of ERVWE2 reference sequence was performed. Several putative binding sites for cellular cofactors and enhancers were found, suggesting that transcription may occur via alternative promoters. However, ERVWE2 DNA sequencing of MS and healthy individuals revealed that all of them harbor a stop codon at site 39, undermining the expression of a full-length protein. Finally, since plaque formation in central nervous system (CNS) of MS patients is attributed to immunological mechanisms triggered by autoimmune attack against myelin, we also investigated the level of similarity between envelope protein and myelin oligodendrocyte glycoprotein (MOG). Comparison of the MOG to the envelope identified five retroviral regions similar to the Ig-like domain of MOG. Interestingly, one of them includes T and B cell epitopes, capable to induce T effector functions and circulating Abs in rats. In sum, although no DNA substitutions that would link ERVWE2 to the MS pathogeny was found, the similarity between the envelope protein to MOG extends the idea that ERVEW2 may be involved on the immunopathogenesis of MS, maybe facilitating the MOG recognizing by the immune system. Although awaiting experimental evidences, the data presented here may expand the scope of the endogenous retroviruses involvement on MS pathogenesis.

Natalizumab treatment for multiple sclerosis: updates and considerations for safer treatment in JCV positive patients.

Natalizumab is currently one of the best options for treatment of patients with Multiple Sclerosis who have failed traditional prior therapies. However, prolonged use, prior immunosuppressive therapy and anti-JCV antibody status have been associated with increased risk of developing progressive multifocal leukoencephalopathy (PML). The evaluation of these conditions has been used to estimate risks of PML in these patients, and distinct (sometimes extreme) approaches are used to avoid the PML onset. At this time, the biggest issue facing the use of Natalizumab is how to get a balance between the risks and the benefits of the treatment. Hence, strategies for monitor JCV-positive patients undergoing Natalizumab treatment are deeply necessary. To illustrate it, we monitored JCV/DNA in blood and urine of a patient receiving Natalizumab for 12 months. We also bring to discussion the effectiveness of the current methods used for risk evaluation, and the real implications of viral reactivation.

New findings about trichodysplasia spinulosa-associated polyomavirus (TSPyV)--novel qPCR detects TSPyV-DNA in blood samples.

A new real-time PCR assay for trichodysplasia spinulosa-associated polyomavirus (TSPyV) DNA detection was designed, and blood samples from kidney transplant recipients and healthy individuals were screened. TSPyV-DNA was not detected in blood from healthy individuals, but 26.8% of kidney recipients presented TSPyV-DNA. This is the first report of TSPyV viremia.

Potential measles transmission risk in mass gatherings: Are we safe for the Olympic games--Rio 2016?

The measles virus, genus Morbillivirus of the Paramyxoviridae family, only infects humans. Measles is transmitted by droplets when the infected individual is coughing or sneezing. It is highly infectious (R0 = 15–20 in a completely susceptible population),1 especially in closed environments such as shopping malls, schools and other confined places. Infected individuals can shed the virus up to 4 days before the appearance of their symptoms and up to 4 days after the onset of symptoms.2 Also, its RNA is still detected in biological samples for many weeks.3 Measles is characterized by its high infectivity. The initial clinical manifestations include runny nose, cough, red and watery eyes and small white spots inside the cheeks. After several days, a maculo-papular exanthema erupts.4 Measles was controlled in some regions, including Brazil. Recently, some outbreaks were reported in the country in the past few years and may represent a risk for transmission during mass gathering events, such as the Olympic Games. For that reason, the aim of this article is to discuss the potential risk for Measles transmission in the country during the Olympic Games and also preventive measures in order to avoid other outbreaks. The number of suspected and confirmed measles cases in 2015 reported by WHO5 indicates a frame, in which the African Region presents the highest number of confirmed cases, whereas Region of Americas presents the lowest: African Region (54 374 suspect/35 557 confirmed), Region of the Americas (13 083 suspect/366 confirmed), Eastern Mediterranean Region (26 178 suspect/11 610 confirmed), European Region (6642 suspect/6023 confirmed), South-East Asia Region (79 713 suspect/67 233 confirmed) and Western Pacific Region (120 319 suspect/61 493 confirmed). This represents a total of 300.309 suspect cases and 182.282 confirmed cases in the world until October 2015. WHO indicates that from June 2014 to June 2015 in the European Continent, there were more …

Pre-transplant shedding of BK virus in urine is unrelated to post-transplant viruria and viremia in kidney transplant recipients

BK virus‐(BKV) associated nephropathy (BKVN) is a major cause of allograft injury in kidney transplant recipients. In such patients, subclinical reactivation of latent BKV infection can occur in the pre‐transplant period. The purpose of this study was to determine whether urinary BKV shedding in the immediate pre‐transplant period is associated with a higher incidence of viruria and viremia during the first year after kidney transplantation. We examined urine samples from 34 kidney transplant recipients, using real‐time quantitative polymerase chain reaction to detect BKV. Urine samples were obtained in the immediate pre‐transplant period and during the first year after transplant on a monthly basis. If BKV viruria was detected, blood samples were collected and screened for BKV viremia. In the immediate pre‐transplant period, we detected BKV viruria in 11 (32.3%) of the 34 recipients. During the first year after transplantation, we detected BKV viruria in all 34 patients and viremia in eight (23.5%). We found no correlation between pre‐transplant viruria and post‐transplant viruria or viremia (p = 0.2). Although reactivation of latent BKV infection in the pre‐transplant period is fairly common among kidney transplant recipients, it is not a risk factor for post‐transplant BKV viruria or viremia.

Lack of evidence of ERVWE1/Xq22.3 involvement in MRSV transcripts detected in Multiple Sclerosis patients

HERV are viral ‘fossils’ that constitute 8% of the human genome and have been implicated in both health and disease conditions. HERV-W/MRSV (Multiple Sclerosis associated RetroVirus) RNA has been detected in patients with multiple sclerosis. MS is a chronic inflammatory disease of the central nervous system of unknown cause, resulting in demyelization and axonal degeneration. MSRV is defined by different transcripts, which vary in their source. Recently, it was described that most of MSRV/Env transcripts from MS patients derived from Xq22.3, 15q21.3 and 6q21. Although the genomic Xq22.3 MRSV/Env is truncated due to a stop codon (TGA) at position 39, the transcripts associated harbored a tryptophan (TGG) instead. Thus, to evaluate if a polymorphism at this position could be involved with full expression of this locus in MS patients we sequenced 1085bp of the ERVWE1/Xq22.3 from 15 MS patients of different ethnic origins and different clinical presentations and 14 health individuals. We found that all MS individuals harbor a stop codon at position 39, undermining the expression of a full-length ENV protein. No additional aminoacid substitution was found in both groups. Also, since there is no LTR flanking this gene, we performed an automated search for promoter sequences in 10kb nearby region. We found several putative binding sites for cellular co-factors and enhancers, suggesting that transcription may occur via alternative promoters. Altogether, the findings suggest that transcripts associated to ERVWE1/Xq22.3 previously detected may be artificially generated, possible due to in vitro recombination between this locus and other active ERVW/Env.

The silent spread of West Nile virus in Brazil: non-human positive case in a beach tourist destination—Espirito Santo

Dear Editor, West Nile virus (WNV) is an emerging arbovirus related to a neurotropic flavivirus that usually infects birds, and can be accidentally transmitted to other animals, including humans. Positive cases in humans are mainly asymptomatic (8 of 10 infected individuals), but a small portion present fever (1 in 5 infected individuals) with headache, myalgia, arthralgia, vomiting, diarrhoea or rash, common symptoms of other flaviviruses. The severe illness, with important neurological consequences, may affect only 1 in 150 of infected patients. Specific diagnostic tests are crucial for surveillance and local control. Evidence of the viral circulation in Brazil was reported in equids in Amazon and Pantanal areas in 2009. The spread to the northeast region of the country, with positive cases in equids and birds, occurred in 2013. One year later, the first human case was detected (Piauí State). WNV was silently transmitted, arriving now in Brazilian’s Southeast Region (Figure 1) according to the report of one equid death in 2018, June in Espírito Santo State, an important domestic tourist destination in Brazil, which currently presents other arboviruses’ outbreaks as Zika (n = 219 reported cases), Chikungunya (n = 482 reported cases) and Dengue (n = 6.945 reported cases). The persistence of other arboviruses in the autumn/winter period points to the vectors population increasing in the region (including Aedes aegypti, Culex quinquefasciatus and others), a similar condition that affects other Brazilian states. Considering that the virus infects birds, equids and also humans, areas with high-population density may be in alert (previous cases occurred in uninhabited locations) due to the fast spread of other arboviruses in the state. The existence of a similar favourable causal network in the USA contributed to the WNV epidemic in 1999. We recommend, especially for domestic travellers, the use of mosquito repellents, especially icaridin, and to wear pants and shirts with long sleeves when visiting natural areas in Espírito Santo State. Unfortunately, there is no cure or vaccine for humans against WNV, only palliative treatment for the symptoms. Other protective measures may be established such as public campaigns for mass information (including preventive instructions to avoid the disease) and the elimination of mosquito breeding sites in the urban areas (to reduce the transmission of arboviruses). Other actions of eco-epidemiological surveillance by the municipal and state government, aiming at fast identification of probable hosts in urban and peri-urban areas may become urgent.

Polyomavirus detection in Multiple Sclerosis patients under Natalizumab therapy: Profile and frequency of urinary shedding

Patients undergoing Natalizumab (NTZ) therapy are at risk of progressive multifocal leukoencephalopathy (PML). Besides John Cunningham virus (JCV), BK polyomavirus might represent an additional concern for such patients since it can also infect CNS cells. Currently, data regarding the presence of anti‐JCV antibodies added to previous immunosuppressive therapy and prolonged NTZ therapy has been used to classify patients at risk of developing PML. Here, we investigated the profile shedding of JCV and BKV in multiple sclerosis (MS) patients during treatment with NTZ. Serial blood and urine samples from 97 MS patients receiving either NTZ or β‐interferon were investigated for polyomavirus shedding. While all blood samples tested negative, 36% of the patients shed polyomavirus in the urine in at least one time point. From these, 21.7%, 9.3%, and 5.1% shed JCV, BKV, and both polyomavirus, respectively. No difference was observed between the rates of urinary shedding of patients treated with NTZ (38.9%) and patients treated with other drugs (34.5%), also no PML event was diagnosed during the follow‐up. Therefore, urinary shedding might not be interfered by therapy condition. In our study, we also observed 14/27 (52%) of anti‐JCV antibodies prevalence, and nearly half of them (42%) did not present any event of urinary shedding during the follow‐up. J. Med. Virol. 89:528–534, 2017.

Human endogenous retrovirus K (HERV-K) transcripts detection in babies exposed to HIV-1 during pregnancy

Human endogenous retroviruses of the K family (HERV-K) are among the most recently integrated retroviruses in the human genome. HERV-K mRNA can be detected in normal tissues, but its expression is remarkably enhanced in HIV-1 infected patients due to HIV-Tat induction. Additionally, HERV-K activity stimulates both humoral and cellular responses, suggesting that HERV-specific immunity may contribute to the control of HIV-1 replication in adults. Recently, it was reported that vertically infected children has HERV specific T-cell response. Since most children born from HIV infected mothers (65-85%) do not get infected despite the possible intra-utero contact with the virus, we evaluated the HERV-K activity in non-infected babies born from: HIV-1 positive mothers (HIV-exposed), and from non-HIV-1 mothers (non-exposed). Peripheral blood mononuclear cells (PBMC) were obtained from 11 babies between 1 to 11-months-old and also from their respective HIV-1 infected mothers. Exposed babies were not breastfed and had no HIV or other viral infection. HERV-K (HML-1-10) transcripts were screened by qRT-PCR, and the expression was evaluated through the absolute threshold-Ct normalized against B-Actin expression. All babies presented some level of HERV-K expression that could not be differentiated between HIV-exposed and non-exposed groups. The expression was also similar according to the age and ethnic origin. Although all HIV-positive mothers have undetectable HIV-1 load, all but one expressed HERV-K. Thus, because all babies investigated presented HERV-K activity despite the HIV-1 exposure, it is possible that HERV activity in babies could works as a primal immunological mechanism to dealing with the exogenous infectious agents.