Share your thoughts on this presentation with #IAS2019

Slides:

Advertisements

Similar presentations

Purging the HIV-1 Reservoir Through the Disruption of the PD-1 Pathway

Advertisements

Measuring the latent HIV Reservoir

CHER Trial: Early Antiretroviral Therapy and Mortality Among HIV- Infected Infants New England J Med 2008;359 (21):

Interactive Workshop “HIV Cure 101”: Challenges in identifying and targeting the HIV reservoir Sarah Palmer Centre for Virus Research Westmead Millennium.

Gag-positive reservoir cells can be identified by a new technique and can be cleared by CTL ? Una O’Doherty June 29, 2013.

2014 “Towards an HIV Cure” symposium Melbourne A novel assay that precisely measures the size of the latent HIV reservoir reveals that ART-nai ̈ ve individuals.

Measuring the latent HIV Reservoir

T-cell activation positively correlates with cell-associated HIV-DNA level in PBMCs in viremic patients with acute or chronic HIV-1 infection Laurence.

KEY CONCEPT Genetics provides a basis for new medical treatments.

Autologous T-cell therapy based on a lentiviral vector expressing long antisense RNA targeted against HIV-1 env gene influences HIV replication and evolution.

Immune correlates of unusual control of viral replication after cessation of HAART Ellen Van Gulck 1, Leo Heyndrickx 1, Céline Merlin 1, Sandra Coppens.

HIV Cellular Pathogenesis III

Evaluation of residual HIV-1 replication among individuals receiving different antiretroviral treatment regimens Giron, LB; Tenore, S; Gabriel, R; Janini,

Viral Tissue Reservoirs Are Determined Early and Little Viral RNA Is Detected during Suppression in the Macaque Model Zandrea Ambrose, Ph.D. Division of.

Anti-HIV Antibody Responses Reflect the Quantifiable HIV Reservoir Size Sulggi Lee, M.D., Ph.D. Assistant Professor of Medicine University of California,

A new molecular mechanism for HIV-1 latency HIV latency in primary T cells: natural activation/purging by DCs Ben Berkhout Academic Medical Center University.

Functional cure after long term HAART initiated during early HIV infection - a case study. van Lunzen J. 1,2, Schulze zur Wiesch J. 1,2, Schumacher U.

LOWERING THE DETECTION LIMITS OF HIV-1 VIRAL LOAD USING REAL-TIME IMMUNO-PCR FOR HIV-1 P24 ANTIGEN Niel T. Constantine, Ph.D., Daniel Edelman, M.S., Janet.

Learning About an HIV Cure by Doing Studies in HIV+ Subjects Alan Landay, PhD Professor and Chairman Department of Immunology/Microbiology Rush University.

Sporadic excesses of unintegrated HIV DNA can occur in HIV+ patients on HAART Una O’Doherty University of Pennsylvania.

KEY CONCEPT Genetics provides a basis for new medical _____________.

Alexander Pasternak Laboratory of Experimental Virology, Department of Medical Microbiology Academic Medical Center of the University of Amsterdam Amsterdam,

The impact of treatment duration on defective and expanded identical HIV genomes in T cell subsets from peripheral blood and tissues Eunok Lee.

Viral and Host characteristics of a child with perinatal HIV-1 after prolonged period of ART cessation in the CHER trial Avy Violari, Mark Cotton, Louise.

Emory University School of Medicine Department of Medicine

Catherine K. Koofhethile 21st IAS July,2016 Durban, SA

In The Name of God.

HIV Cure: Current Status and Future Perspectives

From: Nonthermal Dielectric Barrier Discharge (DBD) Plasma Suppresses Herpes Simplex Virus Type 1 (HSV-1) Replication in Corneal Epithelium Trans. Vis.

Introduction Results Objectives Methods Conclusion Funding

EBV Persistence in Memory B Cells In Vivo

Volume 2, Issue 8, Pages (August 2015)

Detection of bacteria in blood products

V. Bordoni, M. Bibas, I. Abbate, D. Viola, G. Rozera, C. Agrati, A

A Combinatorial CRISPR-Cas9 Attack on HIV-1 DNA Extinguishes All Infectious Provirus in Infected T Cell Cultures Gang Wang, Na Zhao, Ben Berkhout, Atze.

KEY CONCEPT Genetics provides a basis for new medical treatments.

Volume 140, Issue 2, Pages e1 (February 2011)

Volume 20, Issue 3, Pages (September 2016)

Ixazomib reduces HIV-1 reservoir size in a Casp8p41-dependent manner

Evaluation of Candidate Standard XX (97/650)

The microarray technology: facts and controversies

Volume 5, Issue 6, Pages (June 2002)

Volume 21, Issue 3, Pages (October 2017)

Residual HIV replication + virus/cell latency + viral sanctuaries

KEY CONCEPT Genetics provides a basis for new medical treatments.

KEY CONCEPT Genetics provides a basis for new medical treatments.

Volume 154, Issue 6, Pages (May 2018)

Volume 68, Issue 4, Pages (April 2018)

Volume 18, Issue 5, Pages (January 2017)

Andrew J Henderson, Ruth I Connor, Kathryn L Calame Immunity

Fig. 1 HIV transcription profiles in freshly isolated CD4+ T cells and PBMCs from HIV-infected patients on ART suggest blocks to HIV transcriptional elongation,

Figure 2. Technique overview

Volume 158, Issue 5, Pages (August 2014)

KEY CONCEPT Genetics provides a basis for new medical treatments.

Volume 16, Issue 6, Pages (December 2014)

KEY CONCEPT Genetics provides a basis for new medical treatments.

Fig. 2 Activation of freshly isolated CD4+ T cells from HIV-infected patients on ART results in successive increases in elongated, polyadenylated, and.

EBV Persistence in Memory B Cells In Vivo

Introduction. Activation of HIV-Specific CD8+ T Cells from HIV+ Donors by Vesatolimod.

Fig. 3. The effect of VRC01 infusion on virus load in plasma and blood CD4 T cells during ART. The effect of VRC01 infusion on virus load in plasma and.

Improved Detection of the KIT D816V Mutation in Patients with Systemic Mastocytosis Using a Quantitative and Highly Sensitive Real-Time qPCR Assay Thomas.

Triplex-forming Peptide Nucleic Acids Induce Heritable Elevations in Gamma-globin Expression in Hematopoietic Progenitor Cells Joanna Y Chin, Faisal.

Latent human cytomegalovirus enhances HIV-1 infection in CD34+ progenitor cells by Allen Ka Loon Cheung, Yiru Huang, Hau Yee Kwok, Min Chen, and Zhiwei.

Molecular Therapy - Nucleic Acids

Figure 1. Longitudinal measures of HIV persistence and immunologic phenotype/function during anti-PD-1 therapy are ... Figure 1. Longitudinal measures.

KEY CONCEPT Genetics provides a basis for new medical treatments.

Volume 18, Issue 5, Pages (January 2017)

Contribution of naïve CD4+ T cells to the intact HIV reservoir

Share your thoughts on this presentation with #IAS2019

Primary GBM cells support productive HCMV replication in vitro.

Presentation transcript:

Share your thoughts on this presentation with #IAS2019 Highly sensitive, limiting dilution-multiplex seminested quantitative PCR-based assay reveals a large active HIV-1 reservoir in infected ART-suppressed individuals and elite suppressors Alexander Pasternak Laboratory of Experimental Virology, Department of Medical Microbiology, Amsterdam UMC, University of Amsterdam, The Netherlands Share your thoughts on this presentation with #IAS2019

Optional: cell stimulation PBMCs from HIV-infected individual Limiting dilution in 0.5 million uninfected donor PBMC/well RNA isolation by the Boom method DNase treatment One-step RT + 1st PCR (multiplex US + MS RNA) PBMC per dilution 2,000,000 500,000 125,000 31,250 7,813 1,953 488 122 US RNA+ cells MS RNA+ cells qPCR US RNA (gag) qPCR MS RNA (tat/rev/nef) Frequencies of cells with US or MS RNA are determined using the maximum likelihood method Detection limit = 1 HIV RNA+ cell in 7.6 million cells (based on typical nr. of replicates)

All HIV-1 forms detectable in all individuals ** * ** 3 viremic therapy-naïve individuals, 5 aviremic ART-treated individuals, 2 elite suppressors No ex vivo stimulation All HIV-1 forms detectable in all individuals

in integrated DNA+ cells in integrated DNA+ cells % US RNA+ cells in integrated DNA+ cells % MS RNA+ cells in US RNA+ cells % MS RNA+ cells in integrated DNA+ cells

Kevin Groen Maria Feuchert Yara L. Verschoor Marilia R. Pinzone Laura K. DeMaster Una O’Doherty Ben Berkhout a.o.pasternak@amsterdamumc.nl