Simultaneous Assessment of Plasma and Peripheral Mononuclear Cells for Multiple Viral Load Quantification in Peripheral Blood of Patients after Heart Transplantation

2019 ◽  
Vol 38 (4) ◽  
pp. S308-S309 ◽  
Author(s):  
N. Fukushima ◽  
H. Sakaguchi ◽  
K. Toda ◽  
S. Kogaki ◽  
J. Narita ◽  
...  
Keyword(s):  
Viral Load ◽  
Heart Transplantation ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Mononuclear Cells ◽  
Viral Load Quantification ◽  
Simultaneous Assessment

scholarly journals Thyrocytes as the Target Cells for Hhv-6 Infection in Patients with Autoimmune Thyroiditis / Tireocīti Kā Cilvēka Herpesvīrusa-6 Infekcijas Mēríšūnas Pacientiem Ar Autoimūno Tireoidītu

2016 ◽  
Vol 70 (4) ◽  
pp. 160-164
Author(s):  
Alina Sultanova ◽  
Maksims Èistjakovs ◽  
Egils Cunskis ◽  
Katerina Todorova ◽  
Russy Russev ◽  
...  
Keyword(s):  
Viral Load ◽  
Thyroid Gland ◽  
Autoimmune Thyroiditis ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Herpesvirus ◽  
Target Cells ◽  
Tissue Samples ◽  
Gland Tissue ◽  
Thyroid Gland Tissue

Abstract Human herpesvirus-6 (HHV-6) is a ubiquitous betaherpesvirus with immunomodulating properties that have been suggested to play an important role in the development of several autoimmune disorders. Although the primary targets for HHV-6 replication, both in vitro and in vivo, are CD4+ and CD8+ T lymphocytes, some studies have reported the presence of HHV-6 sequences in different solid organs, including in the thyroid gland, showing possible involvement of this herpesvirus in development of autoimmune thyroid disease. The aim of this study was to determine loads of HHV-6 in thyroid gland tissue in comparison to those in peripheral blood of patients with autoimmune thyroiditis. Seven patients [women mean age 45 (28-65)] with histologically confirmed autoimmune thyroiditis were enrolled in this study. Fluorescence-activated cell sorting was used to distinguish and sort lymphocyte populations from peripheral blood mononuclear cells of patients. HHV-6 load was determined by real-time PCR for peripheral blood and thyroid gland tissue samples. Additionally, all results from molecular analyses were compared with histological results obtained by light microscopy. Viral load was detected only in one (46 viral copies/ 1×106cells) blood sample; others were under the detection limit of the used kit. However, in all HHV-6 positive tissue samples viral load was detected in the range of 132-1620 viral copies/106 cells. Substantial HHV-6 load in lymphocyte subpopulations was detected in two of seven patients. HHV-6 load was detected in NK and CD95+ cells of two patients. The obtained results show that thyroid gland cells (tyrocytes) act as target cells for HHV-6.

scholarly journals Identification and Characterization of HIV-1 Latent Viral Reservoirs In Peripheral Blood

2014 ◽  
Vol 53 (1) ◽  
pp. 60-66 ◽  
Author(s):  
Amanda Chargin ◽  
Fangfang Yin ◽  
Min Song ◽  
Srividyabhuvaneswari Subramaniam ◽  
Grace Knutson ◽  
...  
Keyword(s):  
Viral Load ◽  
Viral Replication ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Plasma Viral Load ◽  
Mononuclear Cells ◽  
Clinical Monitoring ◽  
Genotypic Resistance ◽  
Viral Loads ◽  
Hiv 1

Plasma viral load and CD4 counts are effective for clinical monitoring, but they do not give a full representation of HIV-1 quasispecies in cellular reservoirs, the major repository of replication-competent HIV-1 in infected individuals. We sought to develop a diagnostic system that might stimulate the replication-competent HIV-1 reservoirs for enhanced clinical monitoring, including selection of antiretroviral regimens. Whole-blood samples from 45 HIV-infected individuals were collected into 1 ViraStim HIV-1 activation tube and 1 EDTA tube. Samples were tested for viral load and cell type-specific HIV-1 replication. Further, 7 matched activated/nonactivated samples were sequenced using the Trugene HIV-1 genotyping kit. The percentage of patients with replication-competent virus in peripheral blood mononuclear cells (PBMCs) varied, depending on the baseline plasma viral load in the EDTA tubes. Six out of 24 patients with a starting plasma viral load of <20 copies/ml (cp/ml), 6 out of 8 patients with starting viral loads of >20 and <1,000 cp/ml, and 8 out of 13 patients with starting viral loads of >1,000 all showed increases in viral replication of >5-fold. These increases came from cellular reservoirs in blood as determined by simultaneous ultrasensitive subpopulation staining/hybridization in situ (SUSHI). When resistance genotypes in plasma from activation tubes were compared to those from EDTA tubes for 7 patients, all patients showed additional mutations in the activation tube, while 3 patients demonstrated additional genotypic resistance determinants. We show that HIV-1 viral replication can be stimulated directly from infected whole blood. The sequencing results showed that 3 of 7 cases demonstrated additional drug resistance following stimulation.

scholarly journals High Levels of Viral Replication during Primary Simian Immunodeficiency Virus SIVagm Infection Are Rapidly and Strongly Controlled in African Green Monkeys

2000 ◽  
Vol 74 (16) ◽  
pp. 7538-7547 ◽  
Author(s):  
Ousmane Madiagne Diop ◽  
Aïssatou Gueye ◽  
Marisa Dias-Tavares ◽  
Christopher Kornfeld ◽  
Abdourahmane Faye ◽  
...  
Keyword(s):  
Viral Load ◽  
Viral Replication ◽  
Simian Immunodeficiency Virus ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Chronic Phase ◽  
Peripheral Blood Mononuclear ◽  
Viral Loads ◽  
Immunodeficiency Virus ◽  
Green Monkeys

ABSTRACT In contrast to pathogenic human immunodeficiency virus and simian immunodeficiency virus (SIV) infections, chronic SIVagm infections in African green monkeys (AGMs) are characterized by persistently low peripheral and tissue viral loads that correlate with the lack of disease observed in these animals. We report here data on the dynamics of acute SIVagm infection in AGMs that exhibit remarkable similarities with viral replication patterns observed in peripheral blood during the first 2 weeks of pathogenic SIVmac infections. Plasma viremia was evident at day 3 postinfection (p.i.) in AGMs, and rapid viral replication led by days 7 to 10 to peak viremias characterized by high levels of antigenemia (1.2 to 5 ng of p27/ml of plasma), peripheral DNA viral load (104 to 105 DNA copies/106 peripheral blood mononuclear cells [PBMC]), and plasma RNA viral load (2 × 106 to 2 × 108 RNA copies/ml). The lymph node (LN) RNA and DNA viral load patterns were similar to those in blood, with peaks observed between day 7 and day 14. These values in LNs (ranging from 3 × 105 to 3 × 106 RNA copies/106LN cell [LNC] and 103 to 104 DNA copies/106 LNC) were at no time point higher than those observed in the blood. Both in LNs and in blood, rapid and significant decreases were observed in all infected animals after this peak of viral replication. Within 3 to 4 weeks p.i., antigenemia was no longer detectable and peripheral viral loads decreased to values similar to those characteristic of the chronic phase of infection (102to 103 DNA copies/106 PBMC and 2 × 103 to 2 × 105 RNA copies/ml of plasma). In LNs, viral loads declined to 5 × 101 to 103 DNA copies and 104 to 3 × 105 RNA copies per 106 LNC at day 28 p.i. and continued to decrease until day 84 p.i. (<10 to 3 × 104 RNA copies/106 LNC). Despite extensive viremia during primary infection, neither follicular hyperplasia nor CD8+ cell infiltration into LN germinal centers was detected. Altogether, these results indicate that the nonpathogenic outcome of SIVagm infection in its natural host is associated with a rapidly induced control of viral replication in response to SIVagm infection, rather than with a poorly replicating virus or a constitutive host genetic resistance to virus replication.

scholarly journals Factors Associated With Persistence of Plasma HIV-1 RNA During Long-term Continuously Suppressive Firstline Antiretroviral Therapy

2018 ◽  
Vol 5 (2) ◽  
Author(s):  
Alessandra Ruggiero ◽  
Alessandro Cozzi-Lepri ◽  
Apostolos Beloukas ◽  
Douglas Richman ◽  
Saye Khoo ◽  
...  
Keyword(s):  
Viral Load ◽  
Immune Activation ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Viral Load Suppression ◽  
Peripheral Blood Mononuclear ◽  
Treatment Interruptions ◽  
Hla Dr ◽  
Dna 2 ◽  
Hiv 1

Abstract Background Persistence of plasma HIV-1 RNA during seemingly effective antiretroviral thereapy (ART) is incompletely understood. Using an ultrasensitive assay, this cross-sectional study investigated residual plasma HIV-1 RNA in subjects maintained on firstline ART with continuous viral load suppression &lt;50 copies/mL for ≤15 years without recognized viral load blips or treatment interruptions and explored its relationship with the duration of suppressive ART, efavirenz concentrations in plasma, 2-LTR circular HIV-1 DNA (2-LTRc DNA) in peripheral blood mononuclear cells, and cellular (CD4 plus CD26/CD38/CD69; CD8 plus CD38/HLA-DR/DP/DQ) and soluble (sCD14, sCD27, sCD30, IL-6) markers of immune activation in peripheral blood. Methods Residual plasma HIV-1 RNA, total HIV-1 DNA and 2-LTRc DNA were quantified by real-time and digital droplet PCR. Cellular (CD4 plus CD26/CD38/CD69; CD8 plus CD38/HLA-DR/DP/DQ) and soluble (sCD14, sCD27, sCD30, IL-6) markers of immune activation were measured by flow cytometry and ELISA. Results Residual plasma HIV-1 RNA and 2-LTRc DNA were detected in 52/104 (50%) and 24/104 (23%) subjects, respectively. Among subjects with detectable HIV-1 RNA, 50/52 showed levels ≤11 copies/mL. In adjusted analyses, HIV-1 RNA levels were 0.37 log10 copies/mL higher with each log10 U/mL increase in sCD27 (95% confidence interval, 0.01–0.73; P = .02). No significant association was found between residual plasma HIV-1 RNA and other explored parameters. Conclusions These findings point to an ongoing relationship between plasma HIV-1 RNA and selected markers of immune activation during continuously suppressive ART. The novel direct association with levels of sCD27 warrants further investigation.

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