sq-23377 and Viremia

Clonal expansion of HIV-infected cells contributes to the long-term persistence of the HIV reservoir in ART-suppressed individuals. However, the contribution from cell clones that harbor inducible proviruses to plasma viremia is poorly understood. Here, we describe a single-cell approach to simultaneously sequence the TCR, integration sites and proviral genomes from translation-competent reservoir cells, called STIP-Seq. By applying this approach to blood samples from eight participants, we show that the translation-competent reservoir mainly consists of proviruses with short deletions at the 5'-end of the genome, often involving the major splice donor site. TCR and integration site sequencing reveal that cell clones with predicted pathogen-specificity can harbor inducible proviruses integrated into cancer-related genes. Furthermore, we find several matches between proviruses retrieved with STIP-Seq and plasma viruses obtained during ART and upon treatment interruption, suggesting that STIP-Seq can capture clones that are responsible for low-level viremia or viral rebound.

Topics: Anti-Retroviral Agents; CD4-Positive T-Lymphocytes; DNA, Viral; HIV Infections; HIV-1; Humans; Ionomycin; Male; Middle Aged; Phylogeny; Proviruses; Receptors, Antigen, T-Cell; Receptors, Antigen, T-Cell, alpha-beta; Sequence Deletion; Single-Cell Analysis; Viral Load; Viremia

In HIV infection there is a paucity of literature about the degree of immune dysfunction to potentially correlate and/or predict disease progression relative to CD4(+) T cells count or viral load. We assessed functional characteristics of memory T cells subsets as potential prognostic markers for changing viral loads and/or disease progression using the SHIV-infected rhesus macaque model. Relative to long-term non-progressors with low/undetectable viral loads, those with chronic plasma viremia, but clinically healthy, exhibited significantly lower numbers and functional impairment of CD4(+) T cells, but not CD8(+) T cells, in terms of IL-2 production by central memory subset in response to PMA and ionomycine (PMA+I) stimulation. Highly viremic animals showed impaired cytokine-production by all T cells subsets. These results suggest that functional impairment of CD4(+) T cells in general, and of central memory subset in particular, may be a potential indicator/predictor of chronic infection with immune dysfunction, which could be assayed relatively easily using non-specific PMA+I stimulation.

Topics: Animals; Biomarkers; CD4-Positive T-Lymphocytes; Chronic Disease; Cytokines; Disease Progression; Humans; Immunologic Memory; Ionomycin; Macaca mulatta; Prognosis; Simian Acquired Immunodeficiency Syndrome; Tetradecanoylphorbol Acetate; Viral Load; Viremia

Human herpesvirus 8 (HHV-8) is a recently discovered gammaherpesvirus that is the etiologic agent of Kaposi sarcoma (KS). The natural history of primary HHV-8 infection, including clinical outcome and host immune responses that may be important in preventing disease related to HHV-8, has not been elucidated. The present study characterized the clinical, immunologic, and virologic parameters of primary HHV-8 infection in 5 cases detected during a 15-year longitudinal study of 108 human immunodeficiency virus type 1 seronegative men in the Multicenter AIDS Cohort Study. Primary HHV-8 infection was associated with mild, nonspecific signs and symptoms of diarrhea, fatigue, localized rash, and lymphadenopathy. There were no alterations in numbers of CD4(+) or CD8(+) T cells or CD8(+) T-cell interferon gamma (IFN-gamma) production to mitogen or nominal antigen. CD8(+) cytotoxic T-lymphocyte precursor (CTLp) and IFN-gamma reactivity were detected during primary HHV-8 infection, with broad specificity to 5 lytic cycle proteins of HHV-8 encoded by open reading frame 8 (ORF 8; glycoprotein B homolog of Epstein-Barr virus), ORF 22 (gH homolog), ORF 25 (major capsid protein homolog), ORF 26 (a minor capsid protein homolog), or ORF 57 (an early protein homolog), in association with increases in serum antibody titers and appearance of HHV-8 DNA in blood mononuclear cells. CD8(+) T-cell responses to HHV-8 decreased by 2 to 3 years after primary infection. This antiviral T-cell response may control initial HHV-8 infection and prevent development of disease.

Topics: Adult; Amino Acid Sequence; Antibodies, Viral; Antigens, Viral; Capsid; DNA, Viral; Exanthema; Fatigue; Glycoproteins; Herpesviridae Infections; Herpesvirus 8, Human; HIV Seronegativity; Homosexuality; Humans; Immunologic Memory; Immunophenotyping; Incidence; Interferon-gamma; Ionomycin; Longitudinal Studies; Lymphatic Diseases; Lymphocyte Activation; Lymphocyte Count; Male; Middle Aged; Mitogens; Molecular Sequence Data; Phosphoproteins; Prospective Studies; T-Lymphocyte Subsets; Tetradecanoylphorbol Acetate; Viral Envelope Proteins; Viral Matrix Proteins; Viral Proteins; Viremia