The Relationship between Serum Human Immunodeficiency Virus Type 1 (HIV‐1) RNA Level, CD4 Lymphocyte Percent, and Long‐Term Mortality Risk in HIV‐1—Infected Children

ABSTRACT We have been investigating a long-term nonprogressor who was found to be human immunodeficiency virus type 1 (HIV-1) seropositive in 1985 and has survived with stable CD4+ T-cell counts (>1,000 CD4 cells/μl) without any AIDS-related illness. We have previously reported that repeated attempts to measure HIV-1 RNA in the peripheral mononuclear cells obtained from this subject have invariably failed. In the present study, we have analyzed the molecular nature of the HIV-1 quasispecies infecting this patient by PCR amplification of two proviral regions, the 5′ long terminal repeat (5′LTR)/gagleader and the nef gene, directly from fresh uncultured peripheral mononuclear cells, followed by length polymorphism analysis (with 1994, 1995, and 1996 samples) and sequencing (with a 1996 sample). Only proviral forms with nef deletions were revealed by length polymorphism analysis in samples from all three time points. Sequence analysis of the nef gene from the 1996 sample confirmed the presence of similar proviral quasispecies characterized by the presence of several deletions located in thenef-alone and the nef/U3 overlapping regions. Length polymorphism analysis of the 5′LTR/gag leader region suggested the existence of two major quasispecies populations, one characterized by the presence of forms carrying deletions in the U3 region and the other showing a completely intact, full-length 5′LTR. Evidence of the role of nef gene defects in long-term survival of HIV-1-infected patients has been provided so far in two independent investigations involving patients infected with HIV through blood transfusion. Here we show the existence of a similar condition in a subject who acquired HIV-1 seropositivity through the sexual route.

Download Full-text

Effective Suppression of Human Immunodeficiency Virus Type 1 through a Combination of Short- or Long-Hairpin RNAs Targeting Essential Sequences for Retroviral Integration

Journal of Virology ◽

10.1128/jvi.00078-06 ◽

2006 ◽

Vol 80 (15) ◽

pp. 7658-7666 ◽

Cited By ~ 60

Author(s):

Hironori Nishitsuji ◽

Michinori Kohara ◽

Mari Kannagi ◽

Takao Masuda

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Attachment Site ◽

High Dose ◽

Wild Type ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Small interfering RNA (siRNA) could provide a new therapeutic approach to treating human immunodeficiency virus type 1 (HIV-1) infection. For long-term suppression of HIV-1, emergence of siRNA escape variants must be controlled. Here, we constructed lentiviral vectors encoding short-hairpin RNAs (shRNA) corresponding to conserved target sequences within the integrase (int) and the attachment site (att) genes, both of which are essential for HIV-1 integration. Compared to shRNA targeting of the HIV-1 transcription factor tat (shTat), shRNA against int (shIN) or the U3 region of att (shU3) showed a more potent inhibitory effect on HIV-1 replication in human CD4+ T cells. Infection with a high dose of HIV-1 resulted in the emergence of escape mutants during long-term culture. Of note, limited genetic variation was observed in the viruses resistant to shIN. A combination of shINs against wild-type and escape mutant sequences had a negative effect on their antiviral activities, indicating a potentially detrimental effect when administering multiple shRNA targeting the same region to combat HIV-1 variants. The combination of shIN and shU3 att exhibited the strongest anti-HIV-1 activity, as seen by complete abrogation of viral DNA synthesis and viral integration. In addition, a modified long-hairpin RNA spanning the 50 nucleotides in the shIN target region effectively suppressed wild-type and shIN-resistant mutant HIV-1. These results suggest that targeting of incoming viral RNA before proviral DNA formation occurs through the use of nonoverlapping multiple siRNAs is a potent approach to achieving sustained, efficient suppression of highly mutable viruses, such as HIV-1.

Download Full-text

Robust Recall and Long-Term Memory T-Cell Responses Induced by Prime-Boost Regimens with Heterologous Live Viral Vectors Expressing Human Immunodeficiency Virus Type 1 Gag and Env Proteins

Journal of Virology ◽

10.1128/jvi.76.15.7506-7517.2002 ◽

2002 ◽

Vol 76 (15) ◽

pp. 7506-7517 ◽

Cited By ~ 54

Author(s):

Karl Haglund ◽

Ingrid Leiner ◽

Kristen Kerksiek ◽

Linda Buonocore ◽

Eric Pamer ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Vaccinia Virus ◽

Human Immunodeficiency Virus Type ◽

Long Term Memory ◽

Term Memory ◽

Immunodeficiency Virus ◽

High Level ◽

Hiv 1

ABSTRACT We investigated long-term memory and recall cellular immune responses to human immunodeficiency virus type 1 (HIV-1) Env and Gag proteins elicited by recombinant vesicular stomatitis viruses (VSVs) expressing Env and Gag. More than 7 months after a single vaccination with VSV-Env, ∼6% of CD8+ splenocytes stained with major histocompatibility complex class I tetramers containing the Env p18-I10 immunodominant peptide and showed a memory phenotype (CD44Hi). The level of tetramer-positive cells in memory was about 14% of the peak primary response. Recall responses elicited in these mice 5 days after boosting with a heterologous recombinant vaccinia virus expressing HIV-1 Env showed that 40 to 45% of CD8+ splenocytes were tetramer positive and activated (CD62LLo), and these cells produced gamma interferon after stimulation with Env peptide, indicating that they were functional. Five months after the boost, the long-term memory cell population (tetramer positive, CD44Hi) constituted 30% of the CD8+ splenocytes. Recall responses to HIV-1 Gag were examined in mice primed with VSV recombinants expressing HIV-1 Gag protein and boosted with a vaccinia virus recombinant expressing Gag. Using this protocol, we found that ∼40% of CD8+ splenocytes were activated (CD62LLo) and specific for a Gag immunodominant peptide (tetramer positive). The high-level Gag recall response elicited by the vaccinia virus-Gag was greater than that obtained by boosting with a VSV-Gag vector with a different VSV glycoprotein. The corresponding levels of CD44Hi memory cells were also higher long after boosting with vaccinia virus-Gag than after boosting with a glycoprotein exchange VSV-Gag. Our results show that VSV vectors elicit high-level memory CTL responses and that these can be amplified as much as six- to sevenfold using a heterologous boosting vector.

Download Full-text

Long‐Term Evaluation of Cellular Immunity during Antiretroviral Therapy and Immunization with Human Immunodeficiency Virus Type 1 (HIV‐1) Env Glycoprotein in HIV‐1‐Infected Persons

The Journal of Infectious Diseases ◽

10.1086/516500 ◽

1997 ◽

Vol 176 (4) ◽

pp. 904-912 ◽

Cited By ~ 5

Author(s):

Maurizio Carlesimo ◽

Oscar Pontesilli ◽

Emma C. Guerra ◽

Anna R. Varani ◽

M. Livia Bernardi ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Antiretroviral Therapy ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Cellular Immunity ◽

Immunodeficiency Virus ◽

Term Evaluation ◽

Hiv 1

Download Full-text

Dynamics of Human Immunodeficiency Virus Type 1 Replication in Vertically Infected Infants

Journal of Virology ◽

10.1128/jvi.73.1.362-367.1999 ◽

1999 ◽

Vol 73 (1) ◽

pp. 362-367 ◽

Cited By ~ 63

Author(s):

Katherine Luzuriaga ◽

Hulin Wu ◽

Margaret McManus ◽

Paula Britto ◽

William Borkowsky ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Drug Regimen ◽

Turnover Rates ◽

Antiretroviral Drug ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Plasma human immunodeficiency virus type 1 (HIV-1) turnover and kinetics were studied in children aged 15 days to 2 years following the initiation of a triple antiretroviral drug regimen consisting of zidovudine, lamivudine, and nevirapine. HIV-1 turnover was at least as rapid as that previously described in adults; turnover rates were more rapid in infants and children aged 3 months to 2 years than in infants less than 3 months of age. These data confirm the central role of HIV-1 replication in the pathogenesis of vertical HIV-1 infection and reinforce the importance of early, potent combination therapies for the long-term control of HIV-1 replication.

Download Full-text

Comparison of the Quantiplex Version 3.0 Assay and a Sensitized Amplicor Monitor Assay for Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma Samples

Journal of Clinical Microbiology ◽

10.1128/jcm.37.11.3612-3614.1999 ◽

1999 ◽

Vol 37 (11) ◽

pp. 3612-3614 ◽

Cited By ~ 22

Author(s):

Helene C. Highbarger ◽

W. Gregory Alvord ◽

Min Kang Jiang ◽

Akram S. Shah ◽

Julia A. Metcalf ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Rt Pcr ◽

Hiv Rna ◽

Immunodeficiency Virus ◽

Highly Correlated ◽

The Relationship ◽

Hiv 1

This study evaluated correlation and agreement between version 3 of the Quantiplex human immunodeficiency virus type 1 (HIV-1) RNA assay (v3 branched DNA [bDNA]) and a sensitized Amplicor HIV-1 Monitor assay (reverse transcription [RT]-PCR) for the measurement of HIV RNA. Three hundred eighteen samples from 59 randomly selected, HIV-1-seropositive persons on various drug protocols from the National Institute of Allergy and Infectious Diseases HIV outpatient clinic were studied. The results indicate that v3 bDNA and RT-PCR are highly correlated (r = 0.98) and are in good agreement (mean difference in log10 copies/ml ± 2 standard deviations = 0.072 ± 0.371). The relationship between values obtained by both assays is given by the following equation: log10v3 bDNA = −0.0915 + 1.0052 · log10RT-PCR. This represents a 1.026-fold difference between log10RT-PCR values and log10v3 bDNA values.

Download Full-text