Viral Load In Peripheral Blood Research Articles

Management of Cytomegalovirus Infection in Allogeneic Hematopoietic Stem Cell Transplant Recipients

Cytomegalovirus (CMV) is a type of double-stranded deoxyribonucleic acid virus belonging to the herpesviridae family. Following a primary infection, the virus becomes latent in various types of white blood cells. Cytomegalovirus infection can remain latent or become active, especially in immunocompromised individuals, such as those undergoing hematopoietic stem cell transplantation (HSCT), where CMV reactivation can occur. In this context, CMV infection is common and associated with high rates of morbidity and mortality. Pneumonia is one of the most serious complications, with mortality rates exceeding 50%. Additionally, even in the absence of organ-specific disease, CMV infection is related to increased mortality unrelated to hematologic neoplasm recurrence. Given the frequency and severity of this infection in HSCT patients, it is crucial to implement effective strategies for monitoring, prevention, and treatment. This guideline was developed to identify patient groups that benefit from a systematic approach to CMV infection and to define the most appropriate strategy for each group. Monitoring CMV viral load in peripheral blood is crucial, especially in patients at moderate to high risk of active infection. Primary prophylaxis with letermovir (an antiviral drug) is recommended to reduce the incidence of active infection, especially in high-risk patients. Secondary prophylaxis with valganciclovir (antiviral drug) is recommended after an episode of active infection, while preemptive and disease treatment is based on monitoring viral load and clinical response. The aim of this guideline is to improve the approach to CMV infection in HSCT patients, ensuring an effective and safe preventive and therapeutic approach.

Human immunodeficiency virus dynamics in secondary lymphoid tissues and the evolution of cytotoxic T lymphocyte escape mutants.

In secondary lymphoid tissues, human immunodeficiency virus (HIV) can replicate in both the follicular and extrafollicular compartments. Yet, virus is concentrated in the follicular compartment in the absence of antiretroviral therapy, in part due to the lack of cytotoxic T lymphocyte (CTL)-mediated activity there. CTLs home to the extrafollicular compartment, where they can suppress virus load to relatively low levels. We use mathematical models to show that this compartmentalization can explain seemingly counter-intuitive observations. First, it can explain the observed constancy of the viral decline slope during antiviral therapy in the peripheral blood, irrespective of the presence of CTL in Simian Immunodeficiency Virus (SIV)-infected macaques, under the assumption that CTL-mediated lysis significantly contributes to virus suppression. Second, it can account for the relatively long times it takes for CTL escape mutants to emerge during chronic infection even if CTL-mediated lysis is responsible for virus suppression. The reason is the heterogeneity in CTL activity and the consequent heterogeneity in selection pressure between the follicular and extrafollicular compartments. Hence, to understand HIV dynamics more thoroughly, this analysis highlights the importance of measuring virus populations separately in the extrafollicular and follicular compartments rather than using virus load in peripheral blood as an observable; this hides the heterogeneity between compartments that might be responsible for the particular patterns seen in the dynamics and evolution of the HIV in vivo.

Antigen Presenting Cell-Mediated HIV-1 Trans Infection in the Establishment and Maintenance of the Viral Reservoir

Despite potent antiretroviral therapy (ART), an HIV-1 reservoir persists that represents a major barrier to a cure. Understanding the mechanisms by which the HIV-1 reservoir is established and maintained is critical for the discovery of effective treatments to significantly reduce or eliminate the viral reservoir. In addition to cis infection, in which HIV-1 directly infects target CD4+ T cells, cell-to-cell transmission, or trans infection, can also occur. HIV-1 trans infection is significantly more efficient than cis infection, mostly due to the occurrence of multiple infections per cell during transfer. Additionally, trans infection is efficient even in the presence of ART and/or neutralizing antibodies. Cell-to-cell transmission is mediated by CD4+ T cells and professional antigen presenting cells (APC). Here we focus on APC, i.e., myeloid dendritic cells, B lymphocytes, and monocytes/macrophages, that bind, internalize, and transfer HIV-1 to target CD4+ T cells via various proposed mechanisms. We assess the potential impact of trans infection on the establishment and maintenance of the HIV-1 reservoir including its role in disease progression. We consider the natural interactions between APC and CD4+ T cells in vivo that HIV-1 may hijack, allowing for the highly efficient trans infection of CD4+ T cells, maintaining the viral reservoirs in tissue despite undetectable plasma viral loads in peripheral blood. We propose that these modes of viral pathogenesis need to be addressed in potential cure strategies to ensure eradication of the viral reservoir.

Correlation Between CD4 Cell Count, HIV Viral Load, and Chest CT Findings of AIDS-associated Pulmonary Cryptococcosis

Background: The computed tomography (CT) features of acquired immune deficiency syndrome (AIDS)-associated pulmonary cryptococcosis (PC) are correlated with the viral load of human immunodeficiency virus (HIV). An increase in CD4-positive T lymphocyte (CD4) cell count in peripheral blood after a highly active antiretroviral therapy (HAART) can reflect the morphological changes of lung lesions. Objectives: This study aimed to evaluate the correlation between CT features and HIV viral load and to determine a cut-off value for CD4 cell count increment to investigate the prognosis of PC. It also aimed to examine the morphology of PC lesions and their prognosis following HAART. Patients and Methods: Sixty-two patients with AIDS-associated PC, confirmed by pathology or follow-up, were enrolled in this study. The CT findings were recorded and classified as nodular, cavitary, and consolidation groups and their subtypes. Forty HIV patients who had undergone HAART were screened in this study, and the outcomes of lung lesions were recorded in a follow-up of 3 - 6 months. The participants were divided into improvement and progression groups. The correlation analysis and the receiver operator characteristic (ROC) curve analysis were used to examine the correlation between CT morphology and HIV viral load and to determine the cut-off value for CD4 cell count increment. The intraclass correlation coefficient (ICC) for inter-observer agreement was also calculated. Results: In the nodular group, patients with miliary nodules had the highest HIV viral load in peripheral blood (miliary nodules vs. solitary nodules, P = 0.009; miliary nodules vs. multiple nodules; P = 0.024). In the cavitary group, thick-walled cavity lesions had a higher HIV viral load than thin-walled cavity lesions (thin-walled vs. thick-walled cavity lesions, P = 0.036). Changes in the morphology of lesions, indicating the progression or improvement of PC, had a positive correlation with the CD4 cell count increment (F = 4.260, P = 0.045). The cut-off value for CD4 cell count increment to differentiate the two outcomes (progression and improvement) was 44/µL. The area under the curve (AUC) was 0.851, and sensitivity, specificity, and accuracy were estimated at 0.815, 0.714, and 0.764, respectively. Conclusion: In AIDS-associated PC, different types of lesions were related to the HIV viral load, and CD4 cell count increment following HAART was associated with the morphological changes of lesions. This finding can be helpful for clinicians and radiologists to make an accurate diagnosis and evaluate the treatment outcomes, as well as disease progression.

Elevated SARS-CoV-2 in peripheral blood and increased COVID-19 severity in American Indians/Alaska Natives.

Epidemiological data across the United States show health disparities in COVID-19 infection, hospitalization, and mortality by race/ethnicity. While the association between elevated SARS-CoV-2 viral loads (VLs) (i.e. upper respiratory tract (URT) and peripheral blood (PB)) and increased COVID-19 severity has been reported, data remain largely unavailable for some disproportionately impacted racial/ethnic groups, particularly for American Indian or Alaska Native (AI/AN) populations. As such, we determined the relationship between SARS-CoV-2 VL dynamics and disease severity in a diverse cohort of hospitalized patients. Results presented here are for study participants (n = 94, ages 21-88 years) enrolled in a prospective observational study between May and October 2020 who had SARS-CoV-2 viral clades 20A, C, and G. Based on self-reported race/ethnicity and sample size distribution, the cohort was stratified into two groups: (AI/AN, n = 43) and all other races/ethnicities combined (non-AI/AN, n = 51). SARS-CoV-2 VLs were quantified in the URT and PB on days 0-3, 6, 9, and 14. The strongest predictor of severe COVID-19 in the study population was the mean VL in PB (OR = 3.34; P = 2.00 × 10-4). The AI/AN group had the following: (1) comparable co-morbidities and admission laboratory values, yet more severe COVID-19 (OR = 4.81; P = 0.014); (2) a 2.1 longer duration of hospital stay (P = 0.023); and (3) higher initial and cumulative PB VLs during severe disease (P = 0.025). Moreover, self-reported race/ethnicity as AI/AN was the strongest predictor of elevated PB VLs (β = 1.08; P = 6.00 × 10-4) and detection of SARS-CoV-2 in PB (hazard ratio = 3.58; P = 0.004). The findings presented here suggest a strong relationship between PB VL (magnitude and frequency) and severe COVID-19, particularly for the AI/AN group.

Prospective Study of High Dose Foscarnet Prophylaxis for HHV-6 Related CNS Diseases in Cord Blood Transplantation for Adult Patients

Background: Umbilical cord blood transplant (UCBT) recipients showed higher incidence of HHV-6 related CNS diseases (HHV-6 RCD) than those of bone marrow (BM) or peripheral blood (PB). It causes short-term memory impairment or disorientation, which sometimes become irreversible, and convulsions in severe cases, resulted in high non-relapse mortality. We reported previously that in the group given higher doses of prophylactic Foscarnet (FCV) (≥60mg/kg), the incidence of HHV-6 RCD was reduced (Asano-Mori Y, et al. ASH 2013). We thus conducted a prospective study to assess efficacy of higher dose (120mg/kg) of prophylactic FCV to further reduce the incidence and to improve outcome of HHV-6 RCD.Patients and Methods: The study was started since May 2014 and was finished on September 2015. The protocol was approved by local IRB and UCBT recipients who gave written informed consent were included. FCV 120 mg/kg was administered prophylactically from day 7 to day 50 post UCBT.. FCV dose was adjusted according to renal function. The incidence of HHV-6 RCD, types of manifestation and severity of the symptoms, treatment response, and prognosis were evaluated.Results: Among 113 who were included in this study, 9 failed to start FCV from day 7 due to early death or renal impairment, 37 were switched to other drugs at some point before day 50 for reasons such as renal impairment or viral infections other than HHV-6 (23 changed to GCV, 10 to ACV, and 4 to CDV), and 67 received planned dose of FCV. Fifteen developed HHV-6 RCD (12 HHV-6 limbic encephalitis (LE) and 3 myelitis). The cumulative incidence of HHV-6 RCD was 11.5%. No significant risk factors before transplant associated with the development of HHV-6 RCD were identified. Six out of 22 who developed severe pre-engraftment immune reactions (PIR) developed HHV-6 RCD. PIR was shown to be a significant risk factor for HHV-6 RCD development when it was treated as a time-dependent covariate. The median onset day was day 40, and 4 out of 15 patients developed after the cessation of FCV. Only one of 12 encephalitis patients developed convulsions. The median number of copies of HHV-6 in the cerebrospinal fluid at onset was 40,400/µL. HHV-6 viral load in peripheral blood was examined on the same day of CSF examination in 13 and was not detected in 7. Six patients were treated with FCV and GCV in combination, 4 received increased dose of FCV, and 1 was switched from FCV to GCV. Two out of 4 patients who developed after the prophylactic administration of FCV resumed FCV, and the remaining 2 resumed GCV. The virus became undetectable in all 13 evaluable patients by follow-up CSF examination after treatment. The median day of viral disappearance from CSF was 19 days (8-34 days) after diagnosis. All but 2 who were unevaluable due to other complications, had their symptoms improved without sequalae. The median time from treatment intervention to symptomatic improvement was 28 days. The OS after 1 year from HHV6 encephalitis myelitis group was 46.2%, and there was no significant difference in the prognosis as 47.3% OS after 1 year from non-onset group when time-dependent co-analysis was performed (HR 1.88, 95%CI;0.49-2.84, p=0.69). Analysis of OS for 67 patients who were able to undergo preventative FCV administration and other group showed 53.2% and 38.3% respectively in 1 year, revealing a trend in which OS appeared to become higher as a result of continued prophylaxis.Conclusion: There was no difference in the incidence of HHV-6 RCD compared to the previous report even after prophylactic administration of high dose 120 mg/kg of FCV. However, there were few severely ill individuals, most of their symptoms improved due to antiviral treatment without significant sequalae. DisclosuresNo relevant conflicts of interest to declare.

Bimodal distribution and set point HBV DNA viral loads in chronic infection: retrospective analysis of cohorts from the UK and South Africa.

Hepatitis B virus (HBV) viral load (VL) is used as a biomarker to assess risk of disease progression, and to determine eligibility for treatment. While there is a well recognised association between VL and the expression of the viral e-antigen protein, the distributions of VL at a population level are not well described. We here present cross-sectional, observational HBV VL data from two large population cohorts in the UK and in South Africa, demonstrating a consistent bimodal distribution. The right skewed distribution and low median viral loads are different from the left-skew and higher viraemia in seen in HIV and hepatitis C virus (HCV) cohorts in the same settings. Using longitudinal data, we present evidence for a stable 'set-point' VL in peripheral blood during chronic HBV infection. These results are important to underpin improved understanding of HBV biology, to inform approaches to viral sequencing, and to plan public health interventions.

Bimodal distribution and set point HBV DNA viral loads in chronic infection: retrospective analysis of cohorts from the UK and South Africa

Hepatitis B virus (HBV) viral load (VL) is used as a biomarker to assess risk of disease progression, and to determine eligibility for treatment. While there is a well recognised association between VL and the expression of the viral e-antigen protein, the distributions of VL at a population level are not well described. We here present cross-sectional, observational HBV VL data from two large population cohorts in the UK and in South Africa, demonstrating a consistent bimodal distribution. The right skewed distribution and low median viral loads are different from the left-skew and higher viraemia in seen in HIV and hepatitis C virus (HCV) cohorts in the same settings. Using longitudinal data, we present evidence for a stable ‘set-point’ VL in peripheral blood during chronic HBV infection. These results are important to underpin improved understanding of HBV biology, to inform approaches to viral sequencing, and to plan public health interventions.

Chronic Hepatitis B Infection Is Associated with Increased Molecular Degree of Inflammatory Perturbation in Peripheral Blood.

Hepatitis B virus (HBV) infection remains a major public health concern. The interaction between HBV and the host inflammatory response is an important contributing factor driving liver damage and diseases outcomes. Here, we performed a retrospective analysis employing an adapted molecular degree of perturbation (MDP) score system to assess the overall inflammatory imbalance related to persistent HBV infection. Plasma levels of several cytokines, chemokines, and other inflammatory markers were measured in Brazilian individuals diagnosed with either chronic HBV or previous HBV infection, as well as in uninfected controls between 2006 and 2007. Multidimensional analyses were used to depict and compare the overall expression profile of inflammatory markers between distinct clinical groups. Chronic HBV patients exhibited a marked inflammatory imbalance, characterized by heightened MDP scores and a distinct profile of correlation networks inputting plasma concentrations of the biomarkers, compared with either individuals with previous HBV or controls. Furthermore, in participants with chronic HBV infection, the viral loads in peripheral blood were directly proportional to overall molecular perturbation as well as to specific perturbations of interleukin (IL)-4 and interferon (IFN)-γ concentrations. These findings highlight additional nuances about systemic inflammation related to persistent HBV infection.

Feasible alternatives to DBS in the retrospective diagnosis of congenital cytomegalovirus infection

Retrospective diagnosis of congenital cytomegalovirus (cCMV) infection may be challenging mainly because of the high variable sensitivity of PCR on dried blood spots (DBS) samples. To compare cytomegalovirus (CMV) viral load (VL) values in different samples obtained at birth from infants with cCMV infection. To evaluate dried umbilical cord (DUC) samples as an alternative to DBS. Saliva and/or urine, peripheral blood (PB), and DBS from 16 infants with confirmed cCMV infection were collected at birth. CMV VL were determined by DNA extraction and real-time polymerase chain reaction (rt-PCR). In two cases, VL was determined from DUC samples. Six (37.5 %) of the 16 infants were symptomatic, and 10 (62.5 %) were asymptomatic. The CMV VL found in saliva (median: 1,958,525 [IQR: 597,683-3,483,843] IU/mL) and in urine (median: 691,865 [IQR: 188,489.5-3,175,696] UI/mL) were both higher than those found in PB (median: 1115 [IQR: 364-4,002] IU/mL), p: 0.0001). Symptomatic infants presented 100 % of detectable VL in PB and 50 % in DBS. Asymptomatic infants showed 75 % of detectable VL in PB and 30 % in DBS. The VL in DUC were 22,341, 9754 IU/mL and 994 IU/mL. When VL was detectable in PB, the values were lower than in saliva or urine, in both symptomatic and asymptomatic cases of cCMV. The low sensitivity in DBS samples could be due to low blood volume content, making CMV VL undetectable even when using optimised extraction and PCR protocols. In our limited experience, DUC could play a complementary diagnostic role when DBS VL is undetectable.

Bimodal distribution and set point HBV DNA viral loads in chronic infection: retrospective analysis of cohorts from the UK and South Africa

Hepatitis B virus (HBV) viral load (VL) is used as a biomarker to assess risk of disease progression, and to determine eligibility for treatment. While there is a well recognised association between VL and the expression of the viral e-antigen (HBeAg) protein, the precise determinants and distribution of VL at a population level are not well described. We here report the distribution of HBV VL in two large cross-sectional population cohorts in the UK and in South Africa, demonstrating a consistent bimodal distribution. The right skewed distribution and low median viral loads are significantly different from the left-skew and higher viraemia in seen in comparable HIV and hepatitis C virus (HCV) cohorts. Using longitudinal data, we present evidence for a stable 'set-point' VL in peripheral blood during chronic HBV infection. These results are important to underpin improved understanding of HBV biology and to plan public health interventions.

Revealing uninfected and infected target cell dynamics from peripheral blood data in highly and less pathogenic simian/human immunodeficiency virus infected Rhesus macaque

Since chimeric simian and human immunodeficiency viruses (SHIVs) used here, that is, SHIV-#64 and -KS661 utilize both CCR5 and CXCR4 chemokine receptors, they have broad target cell properties. A highly pathogenic SHIV strain, SHIV-KS661, causes an infection that systemically depletes the CD4+ T cells of Rhesus macaques, while a less pathogenic strain, SHIV-#64, does not cause severe symptoms in the macaques. In our previous studies, we established in vitro quantification system for virus infection dynamics, and concluded that SHIV-KS661 effectively produces infectious virions compared with SHIV-#64 in the HSC-F cell culture. However, in vivo dynamics of SHIV infection have not been well understood. To quantify SHIV-#64 and -KS661 infection dynamics in Rhesus macaques, we developed a novel approach and analyzed total CD4+ T cells and viral load in peripheral blood, and reproduced the expected dynamics for the uninfected and infected CD4+ T cells in silico. Using our previous cell culture experimental datasets, we revealed that an infection rate constant is different between SHIV-#64 and -KS661, but the viral production rate and the death rate are similar for the both strains. Thus, here, we assumed these relations in our in vivo data and carried out the data fitting. We performed Bayesian estimation for the whole dataset using MCMC sampling, and simultaneously fitted our novel model to total CD4+ T cells and viral load of SHIV-#64 and -KS661 infection. Our analyses explained that the Malthusian parameter (i.e., fitness of virus infection) and the basic reproduction number (i.e., potential of virus infection) for SHIV-KS661 are significantly higher than those of SHIV-#64. In addition, we demonstrated that the number of uninfected CD4+ T cells in SHIV-KS661 infected Rhesus macaques decreases to the significantly lower value than that before the inoculation several days earlier compared with SHIV-#64 infection. Taken together, the differences between SHIV-#64 and -KS661 infection before the peak viral load might determine the subsequent destiny, that is, whether the systemic CD4+ T cell depletion occurs or the host immune response develop.

Suppression of Active HIV-1 Infection in CD34+ Hematopoietic Humanized NSG Mice by a Combination of Combined Antiretroviral Therapy and CCR5 Targeting Drugs.

Significant progress has been made in the diagnostics and treatment of AIDS since the discovery of the human immunodeficiency virus type 1 (HIV-1) in 1983. The remarkable effectiveness of combined antiretroviral therapy (cART) is evidenced by mortality reduction, control of peripheral blood viral load, and in a nearly normal quality of HIV patients' lives. Remaining obstacles in treatment and cure are drug toxicities and side effects, viral resistance, persistence of HIV-1 reservoirs on termination of cART treatment, the cost of lifelong antiretroviral therapy, and the stigma associated with taking antiretroviral drugs. As determined by plasma viral RNA and peripheral blood mononuclear cells (PBMC) proviral DNA, we show improved suppression of productive HIV infection in human CD34+ hematopoietic stem cell-engrafted NOD (nonobese diabetic)-SCID (severe combined immunodeficiency)-il2rg-/- (NSG) mice by combined treatment with cART and CCR5 targeting drugs, compared with cART alone, as well as an increased preservation of human CD4+ T cells (defined as CD45+ CD3+ CD4+ cells) and CD4+/CD8+ cell ratios in infected mice. The data also suggest a possible reduction in viral reservoirs. Our data confirm that this animal model is suitable for detection of productive HIV infection, replication, and establishment of viral reservoirs. The data also provide proof of principle for the utility of combining CCR5 targeting drugs, maraviroc and rapamycin, with traditional cART to improve control of viremia and reduce viral reservoirs. This study thus serves as a model for future HIV-1 studies that could lead to the clinical development of new generations of antiretroviral drugs.

Management of post-transplant lymphoproliferative disorders.

Management of post-transplant lymphoproliferative disorders.

Diagnostics as Prevention - A Rapid Testing-Based Strategy of Sex Workers against Sexual HIV Exposure.

IntroductionGerman sex workers have illegally established a prevention strategy, which consists of testing potential sexual partners with human immunodeficiency virus (HIV)-specific rapid diagnostic tests (RDTs) prior to engaging in unprotected sexual intercourse eventually performed in case of a negative test result. Based on a recently established modeling approach, the effectiveness of this strategy regarding the risk of HIV exposure was compared with protection provided by condom use.MethodsBased on a literature search, the following assumptions were used for the calculations: an averaged 80% exposure risk reduction with a condom used during sexual intercourse, usage of a well-characterized 4th-generation HIV RDT, and a 10 day post-infection period without any measurable viral load in peripheral blood followed by a sero-conversion period of about 3 weeks with 12.3% test sensitivity (antigen-specific) and only afterwards 97.3% (antibody-specific) test sensitivity.ResultsIn most constellations, the HIV exposure risk in case of RDT-based prevention was lower than with condom use. Conclusions: The RDT-based HIV exposure prevention as established by sex workers is effective in most situations. A notable weakness of the strategy is the RDTs' poor sensitivity in spite of a high transmission risk during the seroconversion stage.

Epidemiology and outcome of chronic high Epstein-Barr viral load carriage in pediatric kidney transplant recipients.

The development of EBV infection and PTLD is normally associated with a high EBV viral load in peripheral blood. Observations have previously identified existence of a CHL carrier state that demonstrated variable outcomes based upon the organ which was transplanted. Data defining the incidence and outcome of CHL in pediatric KTx are not well described. The charts of children undergoing isolated KTx at Children's Hospital of Pittsburgh between January 2000 and December 2014 were retrospectively reviewed. EBV loads in the peripheral blood were routinely measured as part of surveillance protocols at our center. CHL was defined as the presence of high load for >50% of samples for ≥6months. PTLD was defined histologically using WHO definitions. Of 188 isolated KTx recipients, we identified a total of 16 (8%) children who developed CHL carrier state. No patient developed EBV-driven late-onset PTLD. Age at the time of KTx was significantly lower in the CHL group (median 3.9years, interquartile range: IQR 2.9-6.6, P=.0004). Children in the CHL group were more likely to be EBV-seronegative prior to KTx (94%, 15/16), compared to the UVL and LVL groups (55% and 50%, respectively, P<.002). The median duration of CHL carrier state was 20months (IQR 10.7-35.8). Fifteen of the 16 CHL carriers experienced spontaneous resolution of CHL carrier state. Children in the CHL group were younger at the time of primary EBV infection (P=.023). Finally, antiviral medication was not effective in either preventing or decreasing the EBV viral load in blood (P=.84). Overall incidence of late-onset PTLD is very low compared to heart and intestinal transplant, even though KTx recipients can develop CHL carrier state. The CHL carriers in KTx recipients were EBV-seronegative prior to transplant and were younger both at the time of KTx and at the time of primary EBV infection compared to those in the UVL and HVL groups. Antivirals did not prevent EBV infection or decrease EBV viral loads.

Plasma magnesium is inversely associated with Epstein-Barr virus load in peripheral blood and Burkitt lymphoma in Uganda

BackgroundEpstein-Barr virus (EBV) causes endemic Burkitt lymphoma (eBL). EBV control was improved by magnesium (Mg2+) supplementation in XMEN, an X-linked genetic disease associated with Mg2+ deficiency, high circulating EBV levels (viral loads), and EBV-related lymphomas. We, therefore, investigated the relationship between Mg2+ levels and EBV levels and eBL in Uganda. MethodsPlasma Mg2+ was measured in 45 women with low or high circulating EBV levels, 40 pediatric eBL cases, and 79 healthy children. Mg2+ uptake by T-lymphocytes was evaluated in samples from healthy donors. ResultsPlasma Mg2+ deficiency (plasma level <1.8 mg/dl) was more likely in women with high- vs. low-EBV levels (76.0% vs. 35%; odds ratio [OR] 11.3, 95% CI 2.14–60.2), controlling for age, and in eBL cases than controls (42.0% vs. 13.9%; OR 3.61, 95% CI 1.32–9.88), controlling for sex, age group, and malaria status. Mg2+ uptake by T-lymphocytes was related to extracellular Mg2+ concentration. InterpretationPlasma Mg2+ deficiency is associated with high EBV levels and eBL.

Torque Teno Virus (TTV) and Epstein–Barr Virus (EBV) Viral Load in Peripheral Blood as a Biomarker of “Net Immunosuppression”: Factors Influencing Viral Load in Adult Patients Awaiting Solid Organ Transplantation

BackgroundQuantitative measurement of DNA levels of persistent highly prevalent viral infections such as Torque Teno Virus (TTV) and Epstein–Barr Virus (EBV) that are highly dependent on T-cell responses for control have been proposed as possible biomarkers of “net immunosuppression.”MethodsWe examined factors influencing TTV in 51 adult patients awaiting solid organ transplantation (15 kidney, 15 liver/small bowel, 10 heart, 11 lung) and 51 healthy age- and sex-matched controls. Samples from all patients were tested for TTV DNA (plasma) using quantitative nucleic acid testing (QNAT), EBV and cytomegalovirus (CMV) serology, CMV DNA (QNAT of plasma, urine, saliva), and EBV DNA (QNAT of whole blood [WB], saliva). Effects of age, sex, organ, CMV serology, and EBV DNA on TTV DNA were explored in univariate and multivariate models.ResultsPatients and controls were well-matched for age (median 48.4 vs. 49.2 years), sex 64.7% M, both CMV seropositivity (45.1% vs. 51%) and EBV seropositivity (94.1% vs. 98.0%), and detection of EBV DNA in saliva (52.9% vs. 50.0%) and WB (28.6% vs. 16.3%). TTV DNA was detected more frequently and at higher levels (Figure 1) in patients (86.3%) vs. controls (66.7%). TTV DNA plasma levels were age-dependent for patients but not controls (<50 years vs. >50 years, P = 0.021), and were only higher in patients >50 years vs. controls (P = 0.003). Only two patients and no controls had CMV DNA detected. Neither CMV serostatus nor sex impacted TTV DNA levels in patients or controls. Patients awaiting lung transplant had lower TTV levels compared with those awaiting liver (P = 0.04) or heart transplant (P = 0.03). A significant correlation between plasma TTV DNA levels and WB EBV DNA (r = 0.28, P = 0.05) but not saliva EBV DNA levels was observed in patients (Figure 2); only WB EBV DNA level was a significant predictor of high (≥3rd quartile of control) TTV DNA levels in patients in multivariate analysis (P = 0.026). Four of the five patients tested before and 27–285 days after transplant had a significant post-transplant rise in TTV DNA levels (Figure 3).ConclusionMeasurement of TTV DNA in plasma alone or combined with EBV DNA in WB may be a useful marker for measuring global immunosuppression including age-related immunosenescence in subjects not receiving exogenous immunosuppression.Disclosures All authors: No reported disclosures.

High Epstein-Barr Virus Load and Genomic Diversity Are Associated with Generation of gp350-Specific Neutralizing Antibodies following Acute Infectious Mononucleosis.

Antibodies against viral surface proteins can blunt the spread of viral infection by coating viral particles, mediating uptake by immune cells, or blocking interaction with host cell receptors, making them a desirable component of a sterilizing vaccine. The EBV surface protein gp350 is a major target for antibodies. We report the detection of EBV gp350-specific antibodies capable of neutralizing EBV infection in vitro The majority of gp350-directed vaccines focus on glycoproteins from lab-adapted strains, which may poorly reflect primary viral envelope diversity. We report some of the first primary gp350 sequences, noting that the gp350 host receptor binding site is remarkably stable across patients and time. However, changes in overall gene diversity were detectable during infection. Patients with higher peripheral blood viral loads in primary infection and greater changes in viral diversity generated more efficient antibodies. Our findings provide insight into the generation of functional antibodies, necessary for vaccine development.

Possible Relation of Roseolovirus Infection with Fibromyalgia / Iespējamā Roseolovīrusu Infekcijas Saistība Ar Fibromialīiju

Abstract Fibromyalgia (FM) is a chronic widespread pain disorder that impacts 0.5%-7% of the general population worldwide. The aetiology and pathogenesis of the disease are still unknown. Human herpesvirus-6 and -7 belong to the family Herpesviridae, subfamily Betaherpesvirinae, and genus Roseolovirus and are immunomodulating viruses potentially pathogenic to the nervous system. Presence of anti-HHV-6 and -HHV-7 antibodies and viral genomic sequences, viral loads, HHV-6 variant-specificity, and TNF-α level were studied in 41 FM patients and 50 healthy individuals using polymerase chain reactions, restriction endonuclease analysis and ELISA. There was no difference in the presence of anti-HHV-6 and anti-HHV-7 IgG class antibodies between FM patients and control group individuals. Viral sequences were found in 80.5% of FM patients and in 62.0% of controls. Significantly higher rate of concurrent HHV-6 and HHV-7 infection and higher viral loads in peripheral blood were detected in FM patients compared to the control group individuals. Plasma viremia was detected only in FM patients. Significantly higher TNF-α levels were detected in virus positive FM patients. From all positive cases only in two FM patients HHV-6A was revealed. Significantly higher detection frequency of concurrent HHV-6 and HHV-7 infection, simultaneous HHV-6 and HHV-7 activation, higher viral loads and TNF-α expression levels in primary FM patients than in control group individuals indicate the potential involvement of Roseoloviruses in development of this disorder.