Single-copy assay quantification of HIV-1 RNA in paired cerebrospinal fluid and plasma samples from elite controllers

The source of human immunodeficiency virus type 1 (HIV-1) RNA in cerebrospinal fluid (CSF) during HIV-1 infection is uncertain. The sequence heterogeneity of HIV-1 RNA in simultaneous CSF and plasma samples was characterized for five patients at the baseline and during the first week of antiretroviral therapy by two commercial genotyping methodologies. In individual subjects, the sequences in CSF samples differed significantly from those in plasma. In contrast, the viral sequences in CSF at the baseline did not differ from the sequences in CSF during treatment. Similarly, viral sequences in plasma did not vary over this interval. This study provides evidence that HIV-1 RNA in CSF and plasma arise from distinct compartments.

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Automated Multireplicate Quantification of Persistent HIV-1 Viremia in Individuals on Antiretroviral Therapy

Journal of Clinical Microbiology ◽

10.1128/jcm.01442-20 ◽

2020 ◽

Vol 58 (12) ◽

Cited By ~ 3

Author(s):

Jana L. Jacobs ◽

Melissa A. Tosiano ◽

Dianna L. Koontz ◽

Brittany Staines ◽

Andrew Worlock ◽

...

Keyword(s):

Antiretroviral Therapy ◽

Single Copy ◽

Plasma Samples ◽

Limit Of Quantification ◽

Rna Detection ◽

Low Level ◽

Persistent Viremia ◽

Manual Methods ◽

Automated Platform ◽

Hiv 1

ABSTRACT Clearance of low-level viremia that persists in most HIV-1-positive individuals on antiretroviral therapy (ART) is an important milestone for efforts to cure HIV-1 infection. The level of persistent viremia on ART is generally below the lower limit of quantification (LOQ) of current FDA-cleared plasma HIV-1 RNA assays (20 to 40 copies/ml) but can be quantified by reverse transcriptase PCR (RT-PCR) assays with single-copy sensitivity. Such assays require multistep manual methods, and their low throughput limits the capacity to monitor the effects of interventions on persistent viremia. Recently, S. Bakkour, X. Deng, P. Bacchetti, E. Grebe, et al. (J Clin Microbiol 58:e01400-20, 2020, https://doi.org/10.1128/JCM.01400-20), reported the use of multiple replicates and Poisson statistics to infer HIV-1 RNA concentrations below the commercial LOQ of an automated platform (Hologic Panther Aptima). Here, we evaluate the detection and quantitation of low-level viremia using the following two adaptions of the automated platform: a multireplicate strategy (9×) and a concentrated single-replicate strategy in which 5 ml of plasma is concentrated by centrifugation (1×, concentrated). We compare these new methods to a recently reported manual integrase-targeting single-copy assay version 2 (iSCA v2). Using laboratory-generated HIV-1 RNA plasma samples at known concentrations, all three methods had similar sensitivity for HIV-1 RNA detection, although iSCA v2 was most sensitive (95% LOD, 2.3 copies/ml), 9× was marginally less sensitive (95% LOD, 3.0 copies/ml), and 1×, concentrated was least sensitive (95% LOD, 3.9 copies/ml). In contrast, for clinical plasma samples, 9× had greater sensitivity than iSCA v2 (82% of samples were quantifiable compared with 62% of samples by iSCA v2). These results support 9× as an acceptable high-throughput alternative to iSCA v2 for quantifying low-level viremia in individuals on ART.

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Kinetics of HIV-1 in cerebrospinal fluid and plasma in cryptococcal meningitis

Infectious Disease Reports ◽

10.4081/idr.2012.e30 ◽

2012 ◽

Vol 4 (1) ◽

pp. 30 ◽

Cited By ~ 1

Author(s):

Diego M. Cecchini ◽

Ana M. Cañizal ◽

Haroldo Rojas ◽

Alicia Arechavala ◽

Ricardo Negroni ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Viral Load ◽

Cryptococcal Meningitis ◽

Clinical Course ◽

Plasma Concentrations ◽

Plasma Samples ◽

Viral Kinetics ◽

Opportunistic Disease ◽

Kinetics Of ◽

Hiv 1

In order to determine HIV-1 kinetics in cerebrospinal fluid (CSF) and plasma in patients with cryptococcal meningitis (CM), we undertook a prospective collection of paired CSF/plasma samples from antiretroviral therapy- free HIV-infected patients with CM. Samples were obtained at baseline (S1) and at the second (S2) and third (S3) weeks of antifungal therapy. HIV-1 CSF concentrations were significantly lower in both S2 and S3 with respect to S1. Plasma concentrations remained stable. HIV-1 concentrations were higher in plasma than CSF in all cases. Patients who survived the episode of CM (but not those who died) showed a decrease in CSF viral load, what suggests different viral kinetics of HIV-1 in the CSF according to the clinical course of this opportunistic disease.

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HIV‐1 Chemokine Coreceptor Utilization in Paired Cerebrospinal Fluid and Plasma Samples: A Survey of Subjects with Viremia

The Journal of Infectious Diseases ◽

10.1086/428095 ◽

2005 ◽

Vol 191 (6) ◽

pp. 890-898 ◽

Cited By ~ 36

Author(s):

Serena S. Spudich ◽

Wei Huang ◽

Annelie C. Nilsson ◽

Christos J. Petropoulos ◽

Teri J. Liegler ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Plasma Samples ◽

Hiv 1

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HIV-1C env and gag Variation in the Cerebrospinal Fluid and Plasma of Patients with HIV-Associated Cryptococcal Meningitis in Botswana

Viruses ◽

10.3390/v12121404 ◽

2020 ◽

Vol 12 (12) ◽

pp. 1404

Author(s):

Nametso Kelentse ◽

Sikhulile Moyo ◽

Mompati L. Mogwele ◽

Doreen Ditshwanelo ◽

Baitshepi Mokaleng ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Cryptococcal Meningitis ◽

Cytotoxic T Lymphocyte ◽

Population Based ◽

Plasma Samples ◽

Plasma Virus ◽

Significant Difference ◽

Hiv Eradication ◽

Ctl Escape ◽

Hiv 1

HIV-1 compartmentalization in reservoir sites remains a barrier to complete HIV eradication. It is unclear whether there is variation in HIV-1 env and gag between cerebrospinal fluid (CSF) and plasma of individuals with HIV-associated cryptococcal meningitis (CM). We compared HIV-1 env characteristics and the gag cytotoxic T-lymphocyte (CTL) escape mutations from CSF and plasma samples. Employing population-based Sanger sequencing, we sequenced HIV-1 env from CSF of 25 patients and plasma of 26 patients. For gag, 15 CSF and 21 plasma samples were successfully sequenced. Of these, 18 and 9 were paired env and gag CSF/plasma samples, respectively. There was no statistically significant difference in the proportion of CCR5-using strains in the CSF and plasma, (p = 0.50). Discordant CSF/plasma virus co-receptor use was found in 2/18 pairs (11.1%). The polymorphisms in the HIV-1 V3 loop were concordant between the two compartments. From the HIV-1 gag sequences, three pairs had discordant CTL escape mutations in three different epitopes of the nine analyzed. These findings suggest little variation in the HIV-1 env between plasma and CSF and that the CCR5-using strains predominate in both compartments. HIV-1 gag CTL escape mutations also displayed little variation in CSF and plasma suggesting similar CTL selective pressure.

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The Relationships between HIV-1 Infection, History of Methamphetamine Use Disorder, and Soluble Biomarkers in Blood and Cerebrospinal Fluid

Viruses ◽

10.3390/v13071287 ◽

2021 ◽

Vol 13 (7) ◽

pp. 1287

Author(s):

T. Walter ◽

Jennifer Iudicello ◽

Debra Cookson ◽

Donald Franklin ◽

Bin Tang ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Immune System ◽

Public Health Problem ◽

Control Group ◽

Csf Biomarkers ◽

Immune System Dysfunction ◽

Immune Biomarkers ◽

Plasma Factor ◽

History Of ◽

Hiv 1

Methamphetamine (METH) use disorder is highly prevalent among people with HIV (PWH) and is a significant public health problem. HIV and METH use are each associated with immune system dysfunction; however, the combined effects on the immune system are poorly understood. This cross-sectional project measured soluble immune biomarkers in plasma and cerebrospinal fluid (CSF) collected from a control group, people with a history of a METH use disorder (METH+), PWH with no history of METH use disorder (HIV+), and PWH with a history of METH use disorder (HIV+/METH+). HIV, METH, and immune dysfunction can also be associated with affective and cognitive deficits, so we characterized mood and cognition in our participants. Two factor analyses were performed for the plasma and CSF biomarkers. Plasma IL-8, Ccl2, VEGF, and 8-isoprostane loaded onto one factor that was highest in the HIV+/METH+ group (p < 0.047) reflecting worse inflammation, vascular injury, and oxidative stress. This plasma factor was also negatively correlated with delayed recall (R = −0.49, p = 0.010), which was worst in the HIV+/METH+ group (p = 0.030 compared to the control group). Overall, these data implicate that combined HIV-1 infection and METH use may exacerbate inflammation, leading to worse cognition.

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