The Molecular Characterization of Intestinal Explant HIV Infection Using Polymerase Chain Reaction-Based Techniques

Janocko, Laura; Althouse, Andrew D.; Brand, Rhonda M.; Cranston, Ross D.; McGowan, Ian

doi:10.1089/aid.2015.0165

Cited by 8 publications

(11 citation statements)

References 67 publications

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“…The one-step SIV gag qRT-PCR has proven to be a more sensitive method than p27 ELISA. Our results are in agreement with the findings of Janocko et al 62 , who demonstrated the feasibility of qRT-PCR as a readout of HIV infection 62 . Also in agreement with this report 62 , qRT-PCR did not shorten the time to detect evidence of infection.…”

Section: Discussionsupporting

confidence: 93%

MZC Gel Inhibits SHIV-RT and HSV-2 in Macaque Vaginal Mucosa and SHIV-RT in Rectal Mucosa

Calenda

Villegas

Barnable

et al. 2017

JAIDS Journal of Acquired Immune Deficiency Syndromes

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The Population Council's microbicide gel MZC (also known as PC-1005) containing MIV-150 and zinc acetate dihydrate (ZA) in carrageenan (CG) has shown promise as a broad spectrum microbicide against HIV, HSV and HPV. Previous data show antiviral activity against these viruses in cell-based assays, prevention of vaginal and rectal SHIV-RT infection and reduction of vaginal HSV shedding in rhesus macaques and also excellent antiviral activity against HSV and HPV in murine models. Recently we demonstrated that MZC is safe and effective against SHIV-RT in macaque vaginal explants. Here we established models of ex vivo SHIV-RT/HSV-2 co-infection of vaginal mucosa and SHIV-RT infection of rectal mucosa in macaques (challenge of rectal mucosa with HSV-2 did not result in reproducible tissue infection), evaluated antiviral activity of MZC and compared qPCR and ELISA readouts for monitoring SHIV-RT infection. MZC (at non-toxic dilutions) significantly inhibited SHIV-RT in vaginal and rectal mucosa and HSV-2 in vaginal mucosa when present during viral challenge. Analysis of SHIV-RT infection and MZC activity by one-step SIV gag qRT-PCR and p27 ELISA demonstrated similar virus growth dynamics and MZC activity by both methods and higher sensitivity of qRT-PCR. Our data provide more evidence that MZC is a promising dual compartment multipurpose prevention technology candidate.

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Section: Discussionsupporting

confidence: 93%

MZC Gel Inhibits SHIV-RT and HSV-2 in Macaque Vaginal Mucosa and SHIV-RT in Rectal Mucosa

Calenda

Villegas

Barnable

et al. 2017

JAIDS Journal of Acquired Immune Deficiency Syndromes

View full text Add to dashboard Cite

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“…There are limitations to the explant HIV-1 infectibility assay related to sensitivity in detecting p24 in the early phase and by tissue decay in the late when maximally provoked, the ex vivo colorectal tissue assay may have detectable HIV-1 using both quantitative real-time-PCR and ELISA p24 readouts as early as day 4, 36,38 however, the significance and reproducibility of these early phase p24 readouts using the ex vivo colorectal infection model have not been validated, requiring a caveat on these findings as well. Further development of more sensitive whole tissue assays utilizing well-characterized labeled viral vectors focusing on the first 24-48 h for viral integration and replication are underway and will help clarify the role of seminal products on enhancing clinically relevant mucosal target cells' infectibility.…”

Section: Discussionmentioning

confidence: 99%

Human Semen or Seminal Plasma Does Not Enhance HIV-1_BaL Ex Vivo Infection of Human Colonic Explants

KordyKattayoun

ElliottJulie

TannerKaren

et al. 2018

AIDS Research and Human Retroviruses

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To determine whether human whole semen (WS) and seminal plasma (SP) either previously frozen or freshly acquired altered ex vivo infectibility of human colonic explants or was associated with histology or toxicity changes, which may influence mucosal HIV-1 transmission in vivo. Pooled human semen samples were freshly obtained from study volunteers (never frozen) and from commercial sources (frozen/thawed). Endoscopically acquired rectal biopsies were evaluated for toxicity following titered ex vivo WS/SP exposure by histological grading and by MTT assay. The ex vivo HIV-1 biopsy challenge model was used to evaluate effects of exposure to either previously frozen or freshly acquired WS/SP on HIV infectibility at a range of viral inocula (10-10 TCID). To evaluate the effects at lower viral inocula of HIV-1 (10-10), experiments in the presence or absence of WS/SP were also performed utilizing TZM-bl cells. MTT assays and histological scoring demonstrated no tissue degradation of biopsies when exposed for 2 h to concentrations of 10% or 100% of either fresh or previously frozen WS/SP. Ex vivo biopsy HIV-1 challenge experiments showed no differences in the presence of freshly acquired or previously frozen/thawed WS/SP compared with control; no differences were seen with lower infectious titers on TZM-bl cells. Within the limits of assay sensitivity and variability, these data show no toxicity or significant enhancement of HIV-1 infectibility of human rectal mucosa using the colorectal explant model with either pooled fresh or frozen/thawed nonautologous human semen.

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“…Tissue based PK / pharmacodynamic (PD) assays are increasingly being used to characterize the efficacy of ARV PrEP agents. The candidate ARV agent is added to tissue samples in vitro which are then challenged with virus to determine whether the PrEP agent has ARV activity [19,20] . A modification of this assay is to deliver the drug in vivo and to then acquire colorectal tissue that is subsequently exposed to virus ex vivo [21] .…”

Section: Introductionmentioning

confidence: 99%

The pharmacokinetics, pharmacodynamics, and mucosal responses to maraviroc-containing pre-exposure prophylaxis regimens in MSM

McGowan

Wilkin

Landovitz³

et al. 2019

Aids

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HPTN 069/ACTG A5305 was a study of 48-week oral PrEP regimens in MSM and transgender women A rectal substudy was included to evaluate drug concentrations in rectal compartment vs. blood, gut-associated lymphoid tissue (GALT) responses to four antiretroviral (ARV) PrEP regimens (maraviroc (MVC), MVC + emtricitabine (FTC), MVC + tenofovir disoproxil fumarate (TDF), and TDF + FTC), and to determine whether ARV exposure was associated with ex vivo suppression of HIV infection in colorectal explants. Methods: CCR5 genotype was characterized using PCR. At baseline and at weeks 24, 48, and 49, GALT phenotype was characterized by flow cytometry, rectal biopsies were challenged with HIV-1 BaL , and tissue and plasma pharmacokinetics were measured via mass spectrometry. Results: Exposure to MVC was not associated with increased expression of CD4+/CCR5+ HIV target T-cells. Significant ex vivo viral suppression compared to baseline was seen at Weeks 24 and 48, ranging from 1.4 to 1.8 log 10 for all study regimens except the MVC-alone arm which did not show statistically significant viral suppression at Week 48. Tissue concentrations of tenofovir (TFV), TFV-diphosphate (DP), and FTC were correlated with viral suppression. Conclusions: MVC-containing HIV PrEP regimens did not increase GALT CD4+ T-cell activation or the CD4+/CCR5+ phenotype. No virologic suppression was seen with MVC-alone at Week 48 compared to combination regimens, suggesting MVC monotherapy might be less effective than combination ARV PrEP regimens.

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The Molecular Characterization of Intestinal Explant HIV Infection Using Polymerase Chain Reaction-Based Techniques

Cited by 8 publications

References 67 publications

MZC Gel Inhibits SHIV-RT and HSV-2 in Macaque Vaginal Mucosa and SHIV-RT in Rectal Mucosa

MZC Gel Inhibits SHIV-RT and HSV-2 in Macaque Vaginal Mucosa and SHIV-RT in Rectal Mucosa

Human Semen or Seminal Plasma Does Not Enhance HIV-1_BaL Ex Vivo Infection of Human Colonic Explants

The pharmacokinetics, pharmacodynamics, and mucosal responses to maraviroc-containing pre-exposure prophylaxis regimens in MSM

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The Molecular Characterization of Intestinal Explant HIV Infection Using Polymerase Chain Reaction-Based Techniques

Cited by 8 publications

References 67 publications

MZC Gel Inhibits SHIV-RT and HSV-2 in Macaque Vaginal Mucosa and SHIV-RT in Rectal Mucosa

MZC Gel Inhibits SHIV-RT and HSV-2 in Macaque Vaginal Mucosa and SHIV-RT in Rectal Mucosa

Human Semen or Seminal Plasma Does Not Enhance HIV-1BaL Ex Vivo Infection of Human Colonic Explants

The pharmacokinetics, pharmacodynamics, and mucosal responses to maraviroc-containing pre-exposure prophylaxis regimens in MSM

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Product

Resources

About

Human Semen or Seminal Plasma Does Not Enhance HIV-1_BaL Ex Vivo Infection of Human Colonic Explants