DNA Load Research Articles

Frosted branch angiitis due to cytomegalovirus-associated unmasking immune reconstitution inflammatory syndrome: a case report and literature review

BackgroundCytomegalovirus (CMV) retinitis is a common opportunistic infection in patients with acquired immunodeficiency syndrome. The common funduscopic manifestations are haemorrhagic necrotising variety and granular variety. Frosted branch angiitis (FBA), as a special form, when it occurred after antiretroviral therapy(ART), could possibly be associated with immune reconstitution. We report a case of FBA secondary to CMV infection-associated unmasking immune reconstitution inflammatory syndrome (IRIS).Case presentationA 27-year-old man with human immunodeficiency virus infection developed FBA after 35 days of ART. The left Aqueous humour (AqH) tested positive for CMV DNA, and the patient was diagnosed with CMV retinitis. The degree of intraocular inflammation was reflected by increased levels of interleukin (IL)-6 and IL-8 in AqH. After anti-CMV treatment and continuous ART for several months, his FBA and vision significantly improved. CMV DNA became undetectable in the left AqH, and the IL-6 and IL-8 levels in AqH decreased.ConclusionFBA could be a sign of CMV-associated unmasking IRIS. Anti-CMV treatment alone or combination with steroid treatment may be administered, depending on the changes in CMV DNA load and immunologic profile of AqH.

HIV/HBV coinfection: temporal trends and patient characteristics, Spain, 2002 to 2018

BackgroundRecent and reliable estimates on the prevalence of coinfection with human immunodeficiency virus (HIV) and hepatitis B virus (HBV) in Europe are lacking.AimLeveraged on a study designed to assess HIV/HCV coinfection prevalence, we assessed the prevalence of HIV/HBV coinfection in Spain in 2018 and compared the results with five similar studies performed since 2002.MethodsThis cross-sectional prevalence study was carried out in 43 centres, and patients were selected using simple random sampling. The reference population comprised 40,322 patients and the sample size were 1,690 patients.ResultsThe prevalence of HIV/HBV coinfection in Spain at the end of 2018 was 3.2%. The prevalence in 2002, 2009, 2015, 2016 and 2017 was 4.9%, 3.4%, 3%, 3.9% and 3%, respectively. Among the HIV/HBV-coinfected patients identified in 2018, 16.7% had cirrhosis according to transient elastography and 26.3% tested positive for antibodies against hepatitis D virus. All HIV/HBV-coinfected patients were receiving drugs with activity against HBV, and 97% of those tested for HBV DNA had an HBV DNA load < 80 IU/mL.ConclusionsThe prevalence of HIV/HBV coinfection in Spain remained stable at around 3% for a decade. Our data could facilitate the design of national programmes to control HBV infection and help identify areas of patient management that need improvement.

Next-generation sequencing combined with serological tests based pathogen analysis for a neurocysticercosis patient with a 20-year history:a case report

BackgroundNeurocysticercosis (NCC) is the most common helminthic infection of the central nervous system (CNS) caused by the larval stage of Taenia solium. Accurate and early diagnosis of NCC remains challenging due to its heterogeneous clinical manifestations, neuroimaging deficits, variable sensitivity, and specificity of serological tests. Next-generation sequencing (NGS)-based pathogen analysis in patient’s cerebrospinal fluid (CSF) with NCC infection has recently been reported indicating its diagnostic efficacy. In this case study, we report the diagnosis of a NCC patient with a symptomatic history of over 20 years using NGS analysis and further confirmation of the pathology by immunological tests.Case presentationThis study reports the clinical imaging and immunological features of a patient with a recurrent headache for more than 20 years, which worsened gradually with the symptom of fever for more than 7 years and paroxysmal amaurosis for more than 1 year. By utilizing NGS technique, the pathogen was detected in patient’s CSF, and the presence of Taenia solium-DNA was confirmed by a positive immunological reaction to cysticercus IgG antibody in CSF and serum samples. The symptoms of the patient were alleviated, and the CSF condition was improved substantially after the anti-helminthic treatment.ConclusionsThis study suggests that combining CSF NGS with cysticercus IgG testing may be a highly promising approach for diagnosing the challenging cases of NCC. Further studies are needed to evaluate the parasitic DNA load in patients’ CSF for the diagnosis of disease severity, stage, and monitoring of therapeutic responses.

Association of subgingival Epstein–Barr virus and periodontitis

Background: The Epstein–Barr virus (EBV) is gaining interest as a possible agent in the etiology of periodontitis. Previous studies have shown controversy on whether EBV DNA in the subgingival periodontal pockets is associated with periodontitis. The present study aimed to seek the potential relationship between EBV and periodontitis. Methods: Samples were taken from gingival crevicular fluid using sterile paper points, and data on sociodemographics, oral health, and periodontal health were recorded. This case-control study of 118 participants included 59 subjects with severe periodontitis and 59 control subjects with mild periodontitis. Quantitative real-time PCR was used to determined EBV load. Results: EBV DNA was detected in 37.3% of the case samples and 18.6% of the control samples. There was no significant difference in a load of EBV DNA between severe and mild periodontitis (p>0.05). The observed load of EBV DNA was up to 4.55x10 5 copies/mL. The detected EBV DNA was significantly associated with the plaque index and the oral hygiene index (p<0.05). Conclusions: Although no significant association was found, EBV may play a role in periodontitis. The real-time PCR methods can be used to monitor the EBV load in gingival crevicular fluid.

Establishment of a Prognostic Nomogram for Patients With Locoregionally Advanced Nasopharyngeal Carcinoma Incorporating TNM Stage, Post-Induction Chemotherapy Tumor Volume and Epstein-Barr Virus DNA Load

ObjectivesTo establish and validate an effective nomogram to predict clinical outcomes for patients with locoregionally advanced nasopharyngeal carcinoma (LA-NPC).Materials and MethodsThe clinicopathological parameters and follow-up information of 402 locoregionally advanced NPC patients (training cohort, n = 302; validation cohort, n = 100) were retrospectively enrolled. The nomogram was built with the important prognostic variables identified by Cox regression analysis. Overall survival (OS) and progression-free survival (PFS) were the primary and secondary endpoints, respectively. The predictive power and clinical utility of the nomogram were assessed using the Harrell concordance index (C-index), calibration curve, and decision curve analysis. We compared the eighth staging system model with the nomogram to analyze whether the model could improve the accuracy of prognosisResultsEpstein–Barr virus (EBV) DNA load, the gross tumor volume (GTVnx), and cervical lymph node tumor volume (GTVnd) after induction chemotherapy were the independent predictors of OS and PFS. The calibration curves indicated superb agreement between the nomogram-predicted probabilities and observed actual probabilities of survival. The C-index and area under the receiver operator characteristic curve (AUC) of the nomogram integrating these significant factors and N stage, and TNM stage were higher than those of the eighth TNM system alone. In addition, the decision curve analyses demonstrated the clinical value and higher overall net benefit of the nomogram. High-risk groups identified by the nomogram had significantly poorer OS and PFS than the low-risk group (p < 0.05).ConclusionsThe multidimensional nomogram incorporating TNM stage, EBV DNA load, and tumor volume after induction chemotherapy led to a more precise prognostic prediction and could be helpful for stratifying risk and guiding treatment decisions in locoregionally advanced NPC patients who have undergone induction chemotherapy and concurrent chemoradiation.

Derivation and Validation of a Nomogram for Predicting 90-Day Survival in Patients With HBV-Related Acute-on-Chronic Liver Failure.

Background: Conventional prognostic models do not fully reflect the severity of hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF). This study aimed to establish an effective and convenient nomogram for patients with HBV-related ACLF.Methods: A nomogram was developed based on a retrospective cohort of 1,353 patients treated at the Third Affiliated Hospital of Sun Yat-sen University from January 2010 to June 2016. The predictive accuracy and discriminatory ability of the nomogram were determined by a concordance index (C-index) and calibration curve, and were compared with current scoring systems. The results were validated using an independent retrospective cohort of 669 patients consecutively treated at the same institution from July 2016 to March 2018. This study is registered at ClinicalTrials.gov (NCT03992898).Results: Multivariable analysis of the derivation cohort found that independent predictors of 90-day survival were age, white blood cell (WBC) count, hemoglobin (Hb), aspartate aminotransferase (AST), total bilirubin (TBil), international normalized ratio, serum creatinine (Cr), alpha fetoprotein (AFP), serum sodium (Na), hepatic encephalopathy (HE), pre-existing chronic liver disease(PreLD), and HBV DNA load. All factors were included in the nomogram. The nomogram calibration curve for the probability of 90-day survival indicated that nomogram-based predictions were in good agreement with actual observations. The C-index of the nomogram was 0.790, which was statistically significantly greater than those for the current scoring systems in the derivation cohort (P < 0.001). The results were confirmed in the validation cohort.Conclusions: The proposed nomogram is more accurate in predicting the 90-day survival of patients with HBV-related ACLF than current commonly used methods.

Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis

Bovine babesiosis is economically the most important arthropod-borne disease of cattle worldwide. The most significant damage caused by bovine babesiosis is attributed to Babesia bovis due to its higher pathogenicity. This study aimed to develop a real-time PCR method followed by HRM (high-resolution melting) analysis for the simultaneous detection of B. bovis and B. bigemina, enabling a semi-quantitative analysis of Babesia levels using a single-tube reaction. The HRM was compared with real-time PCR using species-specific hydrolysis probes. The HRM analysis allowed to differentiate both Babesia species and was sensitive in the detection and differentiation of 10% for each Babesia species in the sample. Our results suggest the use of this method to estimate the prevalence of infections by B. bovis or B. bigemina as an alternative to the methods of absolute quantification by real-time PCR since it neither requires precise estimates of the number of DNA loads nor the construction of calibration curves. The simultaneous detection of the two Babesia species can be used to characterise the infection levels in cattle populations from different geographical regions, allowing a better control of these diseases.

Role of highly sensitive nucleic acid amplification testing for plasma cytomegalovirus DNA load in diagnosis of cytomegalovirus gastrointestinal disease among kidney transplant recipients.

High plasma cytomegalovirus (CMV) DNA load is generally associated with CMV tissue-invasive disease in solid organ transplant recipients. However, some tissue-invasive diseases, especially CMV gastrointestinal (GI) disease, have undetectable to very low plasma CMV DNA loads. Highly sensitive nucleic acid amplification testing (NAAT) has been increasingly used to quantify low-level CMV DNA loads. Our primary objective was to investigate the epidemiology of CMV GI disease and evaluate the validity of plasma CMV NAAT for the diagnosis of CMV GI disease in kidney transplant (KT) recipients. We conducted a retrospective study of all KT recipients who developed CMV GI disease from January 2016 to December 2018. Plasma CMV DNA load was measured using real-time PCR. The cut-off value of plasma CMV DNA load for diagnosing and risk factors of CMV GI disease were analyzed. A total 17 (3.4%) cases of CMV GI disease occurred in 494 KT recipients. Fifteen (88%) patients had CMV D + /R + serostatus. Fourteen (82%) patients developed CMV GI disease within 6months after KT. Plasma CMV DNA loads were detectable in all (100%) patients with a median load 11,102 (IQR 2,935-107,160) IU/ml. A plasma CMV DNA load of 4,063IU/ml was established as al cut-off for diagnosing CMV GI disease (AUC 0.74, sensitivity 76.5%, specificity 70%, PPV 68, NPV 78). In multivariate analysis, prolonged cold ischemic time (HR 1.14, 95% CI 1.05-1.23, P=.002) and CMV D + /R - serostatus (HR 9.31, 95% CI 2.12-40.74, P=.003) were associated with CMV GI disease. Using highly sensitive NAAT could potentially assist in the diagnosis of CMV GI disease in a CMV D + /R + serostatus setting. KT recipients with CMV seromismatch and prolonged cold ischemic time are at higher risk of CMV GI disease.

CIRCULATING TUMOR DNA LOAD AND TOTAL METABOLIC TUMOR VOLUME IN DIFFUSE LARGE B CELL LYMPHOMA (DLBCL) PATIENTS TREATED BY R‐CHOP – FROM REMARC, A LYSA STUDY

Background: Circulating tumor DNA (ctDNA) quantification has been shown to predict therapeutic benefit in patients (pts) with DLBCL. We recently investigated the prognostic impact of total metabolic tumor volume (TMTV) in DLBCL pts responder to R-CHOP in the REMARC study. In the present ancillary study, we wish to analyze the potential correlation of ctDNA with the outcome and relate these results to functional imaging by PET/CT (TMTV). We also aim to correlate the circulating tumor DNA load with the new score TMTV plus ECOG PS. Patients and Methods: 794 pts were included in the study either at diagnosis (n = 446) or at the end of R-CHOP (n = 347). At the end of R-CHOP, 650 pts achieved a complete response and were randomized to receive 2 years of lenalidomide or placebo. Tumoral cfDNA measurements were performed using a dedicated 36 genes panel with the QiaSeq® technology. CtDNA concentrations were expressed in haploid genome equivalents per mL of plasma (hGE/mL) and calculated by multiplying the mean variant allele frequency (VAF) by the concentration of total cfDNA (Bohers et al. Blood Cancer J 2018).The optimal ctDNA cut-offs for PFS and OS were determined by X-tile analyses and confirmed by a training validation method. Results: A total of 240 pts were analyzed, including 62 non-responder and non-randomized pts (NR-NRP) and 173 responder and randomized pts (REMARC pts). 5 pts had no interpretable ctDNA analysis and were excluded. Among the 235 NR-NRP/REMARC cohort, a ctDNA > 3.74 was present in 14% of pts and was predictive for PFS (HR = 1.68, 95%CI 1.02–2.76, p = 0.0397) and OS (HR = 2.15, 95%CI 1.28-3.61, p = 0.0031). The 3y-PFS for pts with a ctDNA > 3.74 was 47.3% (95%CI: 30%-63%) vs 71.5% (95% CI: 65%-77%) for pts with a ctDNA ≤ 3.74. The 3y-OS for pts with a ctDNA > 3.74 was 56.4% (95%CI: 38%-71%) vs 81.8% (95% CI: 76%-87%) for pts with a ctDNA ≤ 3.74. Among the 173 R-CHOP responder patients (REMARC cohort), the 12 months-relapse free survival since last cycle of R-CHOP for pts with a ctDNA > 3.74 at diagnosis was 61.1% (95%CI: 35%-79%) vs 88.7% (95%CI: 82%-93%) for pts with a ctDNA ≤ 3.74 (p = 0.009). However, a ctDNA > 3.74 was not predictive either for PFS (HR = 1.64, 95%CI: 0.84-3.21, p = 0.14) nor for OS (HR = 1.91 95%CI: 0.93-3.89, p = 0.07). Among the 100 pts with ctDNA and TMTV available, median log ctDNA was found significantly higher in TMTV > 220 (n = 58) vs TMTV < 220 (p = 0.002). However, no significant impact of ctDNA (threshold 3.4) and TMTV(threshold 220) was observed on PFS and OS. Conclusion: Using a routinely targeted sequencing panel for ctDNA measurement, we confirmed the prognostic value of ctDNA concentration to predict outcome in DLBCL patients. For patients responding to R-CHOP, high ctDNA level was significantly associated with high TMTV and early relapse. The research was funded by: BMS Keywords: Diagnostic and Prognostic Biomarkers, Aggressive B-cell non-Hodgkin lymphoma No conflicts of interest pertinent to the abstract.

Mutations in domain V of Mycoplasma pneumoniae 23S rRNA and clinical characteristics of pediatric M. pneumoniae pneumonia in Nanjing, China.

ObjectiveTo investigate the prevalence of mutations in domain V of Mycoplasma pneumoniae (MP) 23S ribosomal RNA (rRNA) and the clinical characteristics of pediatric MP pneumonia (MPP) in Nanjing, China.MethodsDomain V of 23S rRNA was sequenced in MP strains collected from children diagnosed with MPP in Nanjing. Clinical and laboratory data were obtained.ResultsAmong the 276 MP strains, 255 (92.39%) harbored mutations, primarily A2063G in domain V of MP 23S rRNA. When children were stratified according to the presence or absence of mutations, no significant differences were found in sex, age, the MP DNA load at enrollment, lymphocyte counts, pulmonary complications, immunomodulator levels, fever duration, the duration of fever after macrolide therapy, and hospital stay. The prevalence of refractory MPP in the two groups was similar. Children with refractory MPP exhibited higher MP DNA loads than those with non-refractory MPP.ConclusionsDespite the high prevalence of the A2063G mutation in domain V of MP 23S rRNA, mutations were not associated with the clinical characteristics of MPP. The MP DNA load significantly differed between refractory and non-refractory MPP.

PLASMA EPSTEIN-BARR VIRUS (EBV) DNA AS A BIOMARKER FOR DIAGNOSIS OF SYRIAN EBV-POSITIVE BURKITT’S LYMPHOMA

Epstein-Barr virus - positive Burkitt’s Lymphoma is defined as the presence of Epstein-Barr virus (EBV) in tumor cells, the standard way to detect (EBV) in Burkitt’s Lymphoma is in-situ hybridization (ISH) of EBV-encoded small RNA (EBERs) in tumor cells. The present study aimed to evaluate plasma Epstein-Barr virus (EBV) DNA as a noninvasive biomarker for diagnosis and prognosis of EBV-positive Burkitt’s lymphoma. The study included 40 newly diagnosed patients with Burkitt’s lymphoma, ranging in age from 4 to 60 years, and 55 sex and age-matched controls. Forty formalin-fixed paraffin-embedded blocks of Burkitt’s lymphoma tissue samples were used to investigate the EBV by in-situ hybridization detection of the EBERs. Plasma EBV DNA was quantified by real-time quantitative polymerase chain reaction (PCR) for all Burkitt lymphoma patients prior to therapy and for control. The results showed that (22/40, 55%) of Burkitt lymphoma were positive for histological EBER, whereas plasma EBV DNA was detectable (range from 1.2×104 to 4.7×106 copies/mL) in all EBV-positive Burkitt lymphoma samples (22/22). EBV DNA was undetectable in all cases of EBV-negative Burkitt lymphomas (18/18) and all healthy control (55/55). It is worth mentioning that our results demonstrated that the EBV DNA load was significantly high in the EBV-positive BL patients suffering poor prognostic state. In conclusion: Plasma EBV-DNA can be used as a noninvasive biomarker for diagnosis and prognosis of EBV-positive Burkitt’s lymphoma.

Plasma Epstein-Barr virus DNA and risk of nasopharyngeal carcinoma in a prospective seropositive population

ObjectivePlasma Epstein-Barr virus (EBV) DNA is considered a biomarker for nasopharyngeal carcinoma (NPC). However, its long-term role in NPC development is unclear.Materials and methodsA total of 1363 participants seropositive for EBV VCA-IgA and EBNA1-IgA in a community-based NPC screening program in southern China were tested for plasma EBV DNA levels by real-time qPCR between 2008 and 2015. New NPC cases were confirmed by active follow-up approach and linkage to local cancer registry through the end of 2016. Cox proportional hazards regression analysis was performed to calculate the hazard ratios (HRs) for NPC risk with plasma EBV DNA.ResultsThirty patients were newly diagnosed during a median 7.5 years follow-up. NPC incidence increased with the plasma EBV DNA load ranging from 281.46 to 10,074.47 per 100,000 person-years in participants with undetectable and ≥ 1000 copies/ml levels; the corresponding cumulative incidence rates were 1.73 and 50%. Furthermore, plasma EBV DNA loads conferred an independent risk for NPC development after adjustment for other risk factors, with HRs of 7.63 for > 3–999 copies/ml and 39.79 for ≥1000 copies/ml. However, the HRs decreased gradually after excluding NPC cases detected in the first 2 to 3 years and became statistically nonsignificant by excluding cases detected during the first 4 years.ConclusionElevated plasma EBV DNA can predict NPC risk over 3 years. Monitoring plasma EBV DNA can be used as a complementary approach to EBV serological antibody-based screening for NPC.

Association of subgingival Epstein–Barr virus and periodontitis

Background: The Epstein–Barr virus (EBV) is gaining interest as a possible agent in the etiology of periodontitis. Previous studies have shown controversy on whether EBV DNA in the subgingival periodontal pockets is associated with periodontitis. The aim of the present study was to seek the potential relationship between EBV and periodontitis. Methods: Data on socio-demographics, oral health, and periodontal health were recorded, and samples were collected from gingival crevicular fluid, using sterile paper point. This case–control study of 118 participants included 59 subjects with severe periodontitis and 59 control subjects with mild periodontitis. The EBV load was determined by quantitative real-time PCR. Results: EBV DNA was detected in 37.3% of the case samples and in 18.6% of the control samples. There was no significant difference in the load of EBV DNA between severe and mild periodontitis (p&gt;0.05). The observed load of EBV DNA was up to 4.55x105 copies/mL. The detected EBV DNA was significantly associated with the plaque index and the oral hygiene index (all p&lt;0.05). Conclusions: A significant association was not found, but EBV might contribute to periodontitis. Gingival crevicular fluid is useful for monitoring the EBV load by the real-time PCR technique.

Antiviral therapy after curative treatment of hepatocellular carcinoma in patients with chronic hepatitis B infection: Is tenofovir or entecavir preferred?

Antiviral therapy after curative treatment of hepatocellular carcinoma in patients with chronic hepatitis B infection: Is tenofovir or entecavir preferred?

Assessment of the association between cytomegalovirus DNAemia and subsequent acute graft-versus-host disease in allogeneic peripheral blood stem cell transplantation: A multicenter study from the Spanish hematopoietic transplantation and cell therapy group.

The potential role of active CMV infection in promoting acute Graft-versus-Host Disease (aGvHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains a matter of debate. We further addressed this issue conducting a retrospective, observational, multicenter study of 632 patients subjected to allogeneic peripheral blood HSCT at 20 Spanish centers. Monitoring of CMV DNA load in plasma or whole blood was performed by real-time PCR assays. Cumulative incidence of CMV DNAemia was 48.9% (95% CI, 45%-52.9%), of any grade aGvHD, 45.6; 95% (CI, 41.3%-50.1%), and of grade II-IV aGvHD, 30.7 (95% CI, 24.9%-36.4%). Overall, development of CMV DNAemia at any level resulted in an increased risk of subsequent all grade (HR, 1.38; 95% CI, 1.08 - 1.76; P=.009) or grade II-IV (HR, 1.58; 95% CI, 1.22 - 2.06; P=.001) aGvHD. The increased risk of aGvHD linked to prior occurrence of CMV DNAemia was similar to the above when only clinically significant episodes were considered for the analyses (HR for all grade aGvHD, 1.48; 95% CI, 1.13 - 1.91; P=.041, and HR for grade II-IV aGvHD, 1.53; 95% CI. 1.13-1.81; P=.04). The CMV DNA doubling time in blood was comparable overall in episodes of CMV DNAemia whether followed by aGvHD or not. Whether CMV replication is a surrogate risk marker of aGvHD or it is causally involved is an important question to be addressed in future experimental research.

A Prospective Study of Cytomegalovirus-Specific Cell-Mediated Immune Monitoring and Cytomegalovirus Infection in Patients With Active Systemic Lupus Erythematosus Receiving Immunosuppressants.

BackgroundThe effects of cytomegalovirus (CMV)-specific cell-mediated immunity (CMI) on CMV infection in patients with autoimmune diseases receiving immunosuppressants have not been explored.MethodsPatients with active systemic lupus erythematosus (SLE) were preemptively monitored for clinically significant CMV infection (CsCMVI; defined as plasma CMV DNA loads >3 log10 IU/mL). CMV-specific CMI was assessed using an enzyme-linked immunosorbent assay (QuantiFERON-CMV [QF]) before as well as 1 and 3 months after intense immunosuppressive therapy.ResultsThe study included 55 patients with active SLE; patients were a mean age (SD) of 34 (13) years and had a median SLE Disease Activity Index 2000 score (SD) of 14 (8), and 93% were female. Most patients had renal involvement (67%), received methylprednisolone (93%), and were CMV-seropositive (95%). Thirteen (23.6%) patients developed CsCMVI. Among patients with active SLE who were QF-negative (QF–) and QF-positive (QF+) before receiving immunosuppressive therapy, 28.6% and 25% developed CsCMVI, respectively (P = .69). However, 1 month postimmunosuppression, more QF– than QF+ patients developed CsCMVI (44.4% vs 11.8%; P = .03; adjusted hazard ratio, 4.97; 95% CI, 1.07–23.10; P = .04).ConclusionsPatients with active SLE and low CMV-specific T-cell responses could develop CMV infection after receiving immunosuppressants. Further studies should focus on CMV-specific CMI among patients with autoimmune diseases.

Hydroa vacciniforme‐like lymphoproliferative disorder: A clinicopathological, immunohistochemical, and prognostic study of 24 cases in China

Hydroa vacciniforme‐like lymphoproliferative disorder (HV‐LPD) is a rare cutaneous disease associated with Epstein–Barr virus infection. We retrospectively analyzed the clinical presentation, histopathological characteristics, and prognostic study of HV‐LPD in 24 Chinese patients. All patients presented with recurrent papulovesicular and necrotic eruptions on the face, neck, and extremities, with 11 showing systemic symptoms. Twenty patients were diagnosed with HV‐LPD in childhood (age < 18 years) and four in adulthood (age ≥ 18 years). The median age at diagnosis was 8.5 years old (range, 2–50). Histopathology revealed variably dense lymphocyte infiltration throughout the dermis. All cases were strongly positive for CD3 and Epstein–Barr encoding region based on in situ hybridization. Of 18 cases with a T‐cell phenotype, 15 harbored monoclonal rearrangements in T‐cell receptor (TCR) genes. Four cases with a natural killer cell phenotype carried polyclonal rearrangements in TCR genes. Among 24 patients, eight (33.3%) received chemotherapy, two (8.3%) allogeneic hematopoietic stem cell transplantation, and both are currently alive without disease. The median follow‐up period was 24 months (range, 7–120) and 23 patients were available: 15 (62.5%) were alive, and eight (33.3%) had died. Fourteen cases had a relapse of disease and three developed lymphoma within 24 months of diagnosis. The mean survival time of childhood‐onset patients was longer than that of adult‐onset patients (36.4 vs. 20.8 months). In summary, the wide clinical course and representative presentation of cases in our center reflect the pedigree characteristics of HV‐LPD. Allogeneic hematopoietic stem cell transplantation should be a preferred choice for relapse and refractory patients due to the poor effect of chemotherapy. Adult‐onset and high serum EBV DNA loads may indicate an increased risk of aggressive disease in patients with HV‐LPD.

Early Initiation of Antiretroviral Therapy Following In Utero HIV Infection Is Associated With Low Viral Reservoirs but Other Factors Determine Viral Rebound.

BackgroundEarly HIV diagnosis allows combination antiretroviral therapy (cART) initiation in the first days of life following in utero (IU) infection. The impact of early cART initiation on infant viral reservoir size in the setting of high-frequency cART nonadherence is unknown.MethodsPeripheral blood total HIV DNA from 164 early treated (day 0–21 of life) IU HIV-infected South African infants was measured using droplet digital PCR at birth and following suppressive cART. We evaluated the impact of cART initiation timing on HIV reservoir size and decay, and on the risk of subsequent plasma viremia in cART-suppressed infants.ResultsBaseline HIV DNA (median 2.8 log10 copies/million peripheral blood mononuclear cells, range 0.7–4.8) did not correlate with age at cART initiation (0–21 days) but instead with maternal antenatal cART use. In 98 infants with plasma viral suppression on cART, HIV DNA half-life was 28 days. However, the probability of maintenance of plasma aviremia was low (0.46 at 12 months) and not influenced by HIV DNA load. Unexpectedly, longer time to viral suppression was associated with protection against subsequent viral rebound.ConclusionsWith effective prophylaxis against mother-to-child transmission, cART initiation timing in the first 3 weeks of life is not critical to reservoir size.

Abdominal aortic aneurysm and virus infection: A potential causative role for cytomegalovirus infection?

An abdominal aortic aneurysm (AAA) is a multifactorial disease with a variety of genetic and environmental risk factors, but the exact mechanism of AAA formation and progression is still not well understood. The present study investigated the frequency of cytomegalovirus (CMV), Epstein-Barr virus (EBV), and papillomavirus types 6 and 11 (HPV6 and HPV11), their impact on clinical manifestations of cardiovascular diseases, and their possible association with inflammation in patients with AAA and healthy volunteers. Genotyping of CMV UL75, EBV LMP-1, and HPV6 and HPV11 E6 was performed by polymerase chain reaction (PCR), while the viral DNA loads were measured by quantitative real-time PCR (qRT-PCR). Cytokine levels were determined by enzyme-linked immunosorbent assays (ELISAs). The CMV UL75 was detected more frequently in the blood of patients with AAA than in the blood of healthy volunteers (32.7% vs. 6.3%, p < 0.0001). Neither EBV LMP-1 nor HPV6 E6 was found in blood and aortic wall biopsies, while the HPV11 E6 was detected in 36.4% of AAA walls. The CMV infection in patients with AAA was associated with an increased risk of hypertension and coronary artery disease (CAD) (OR 9.057; 95% CI: 1.141 - 71.862, p = 0.037; OR 2.575; 95% CI: 1.002 - 6.615, p = 0.049, respectively). Additionally, CMV-infected patients with AAA had higher TNF-α levels compared with non-infected subjects (p = 0.017). Our findings suggest that CMV infection can stimulate local inflammation in the aorta but is not a direct cause of most abdominal aortic aneurysms. This article is protected by copyright. All rights reserved.

Parvovirus B19 DNA detectable in hearts of patients with dilated cardiomyopathy, but absent or inactive in blood.

AimsParvovirus B19 (B19V) is often assumed to be a cause of dilated cardiomyopathy (DCM), based on the quantification of B19V DNA in endomyocardial biopsies (EMB). Whether the presence of B19V DNA correlates with active infection is still debated. Application of the enzyme endonuclease to blood samples results in degradation of B19V DNA remnants but leaves viral particles intact, which enables differentiation between active and past infection. In this study, the susceptibility to degradation by endonuclease of B19V DNA in blood was compared between DCM patients and a control group of recent B19V infections.Methods and resultsTwenty blood samples from 20 adult patients with DCM, who previously tested positive for B19V DNA in EMB and/or blood, were tested with B19V PCR before and after application of endonuclease to the samples. Six blood samples tested positive for B19V DNA with a mean viral load of 2.3 × 104 IU/mL. In five samples, B19V DNA became undetectable after endonuclease (100% load reduction); in one sample DNA load showed a 23% log load reduction (viral load before endonuclease: 9.1 × 104 IU/mL; after: 6.5 × 103 IU/mL). Presence of cardiac inflammation did not differ between patients with B19V DNAemia (1/4) and patients without B19V DNAemia (6/14) ( P value = 1.0). In all 18 control samples of proven recent B19V infections, DNA remained detectable after application of endonuclease, showing only a mean log load reduction of 2.3% (mean viral load before endonuclease: 8.1 × 1011 IU/mL; after: 8.0 × 1011 IU/mL). Load reduction differed significantly between the DCM group and the control group; indicating the presence of intact viral particles in the control group with proven active infection and the presence of DNA remnants in the DCM group (P value = 0.000).ConclusionDuring recent B19V infection, viral DNA levels in blood were unaffected by endonuclease. In contrast, B19V DNA in blood in patients with DCM became undetectable or strongly reduced after application of endonuclease. Circulating viral DNA in this subset of patients with presumed parvovirus‐associated DCM does not consist of intact viral particles. Viral replicative activity cannot be assumed from demonstrating B19V DNA in cardiac tissue or in blood in DCM patients.