Analysis of alternatively spliced human immunodeficiency virus type-1 mRNA species, one of which encodes a novel TAT-ENV fusion protein

Furtado, Manohar R.; Balachandran, Raghavan; Gupta, Phalguni; Wolinsky, Steven M.

doi:10.1016/0042-6822(91)90773-5

Cited by 57 publications

(65 citation statements)

References 62 publications

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“…Specific amplimers were then detected using DIG-labeled oligoprobes that spanned the 5′ tat splice junction (MF5945, 5′-ATTG-TAAAAAGTGTT-splice-GCTWTCATTGC-3′). The designation K refers to the single code letter for G or C, and W refers to A or T for degenerative bases (25). Tat plasmids were kindly provided by Manohar Furtado (Northwestern University, Chicago, Illiniois, USA) and have been described previously (25).…”

Section: Introductionmentioning

confidence: 99%

“…The designation K refers to the single code letter for G or C, and W refers to A or T for degenerative bases (25). Tat plasmids were kindly provided by Manohar Furtado (Northwestern University, Chicago, Illiniois, USA) and have been described previously (25). Digoxigenin-labeled (DIG-labeled) RNA probes were derived as described previously (7,19).…”

Section: Introductionmentioning

confidence: 99%

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HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death

Brodie

Patterson²,

Lewinsohn

et al. 2000

J. Clin. Invest.

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We have tracked the in vivo migration and have identified in vivo correlates of cytotoxic T-lymphocyte (CTL) activity in HIV-seropositive subjects infused with autologous gene-marked CD8 + HIV-specific CTL. The number of circulating gene-marked CTL ranged from 1.6 to 3.5% shortly after infusion to less than 0.5% 2 weeks later. Gene-marked CTL were present in the lymph node at 4.5-to 11-fold excess and colocalized within parafollicular regions of the lymph node adjacent to cells expressing HIV tat fusion transcripts, a correlate of virus replication. The CTL clones expressed the CCR5 receptor and localized among HIV-infected cells expressing the ligands MIP-1α and MIP-1β, CC-chemokines produced at sites of virus replication. Aggregates of apoptotic cells and cells expressing granzyme-B localized within these same sites. In contrast, lymph node sections from untreated HIV-seropositive subjects, all with significant viral burden (> 50,000 HIV RNA copies/mL plasma), showed no CC-chemokine expression and exhibited only sporadic and randomly distributed cells expressing granzymes and/or apoptotic cells. These studies show that the infused CTL specifically migrate to sites of HIV replication and retain their antigen-specific cytolytic potential. Moreover, these studies provide a methodology that will facilitate studies of both the magnitude and functional phenotype of Ag-specific CD8 + T cells in vivo.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death

Brodie

Patterson²,

Lewinsohn

et al. 2000

J. Clin. Invest.

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“…The positions for the remaining primers and probe were numbered relative to the HIV-1 strain HXB2 (Bukrinsky et al, 1993; GenBank accession number K03455). regions) of the PBMC isolate from week 8 after inoculation and brain tissue from macaque 6 was cloned as described (Furtado et al, 1991). Total DNA was prepared from flesh and fixed brain tissue and PBMCs of the HIV-l-infected macaque as described above.…”

Section: In Vitro Infection Of M Nemestrina Cultured Rnonocytes Withmentioning

confidence: 99%

“…HIV-I envelope gene was amplified using nested primer sets (Table 1 ; outer set, KK 1/3; inner set, KK2/4) and cloned into pAMP (Gibco). Automated sequencing was performed using primer set KK8/40 (Table 1) with a 373A Laser Sequencer (Applied BioSystems) as described (Furtado et al,, 1991). studies were performed blind on specimens without the examiner having knowledge of the HIV-1 infection status of the animals.…”

Section: In Vitro Infection Of M Nemestrina Cultured Rnonocytes Withmentioning

confidence: 99%

“…The amplified product was then detected using the internally conserved 3~P-labelled oligonucleotide probe R (see Table 1) (Bukrinsky et al, 1993). HIV-1 transcriptional activity was assessed by PCR for reverse-transcribed tat mRNA as previously described (Furtado et al, 1991). Preliminary experiments indicated that the sensitivity of the assay was approximately 25 copies of tat RNA using synthetically synthesized material.…”

Section: Detection Of H1v-1 In Macaque Brain By Solution-based Pcr Anmentioning

confidence: 99%

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Infection of Macaca nemestrina brain with human immunodeficiency virus type 1

Frumkin¹,

Patterson

Leverenz³

et al. 1995

Journal of General Virology

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We previously reported that pigtailed macaques (Macaca nemestrina) became infected after intravenous inoculation with the LAI strain of human immunodeficiency virus type 1 (HIV-1), an isolate that replicates in both M. nemestrina lymphocytes and blood-derived monocyte/ macrophages. In the current study we investigated the presence of HIV-1 and pathology in the postmortem brains of four of these macaques. Histopathological findings revealed focal lesions in white matter in the frontal and occipital lobes of one macaque, with myelin loss, nerve fibre loss, and gliosis within these lesions. Semi-quantitative, solution-based PCR revealed HIV-1 DNA in the brains of two of the other macaques. Using slide-based PCR-driven in situ hybridization, we studied these two macaques further and detected intranuclear, circular HIV-1 DNA in vascular endothelia and other non-neuronal brain cells. These findings indicate that M. nemestrina brain can be infected with HIV-1 in vivo and may provide a useful animal model for understanding early HIV-1 brain infection in humans.

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Unintegrated HIV-1 circular 2-LTR proviral DNA as a marker of recently infected cells: Relative effect of recombinant CD4, zidovudine, and saquinavir in vitro

Panther¹,

Coombs²,

Aung³

et al. 1999

J. Med. Virol.

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Unintegrated HIV-1 proviral DNA is one of the earliest detectable forms of HIV-1, and the influence of an antiretroviral drug on its appearance may reflect the efficacy of that agent in preventing infection of new cells. We characterized the dynamics of HIV-1 p24 (p24) antigen production, HIV-1 gag DNA, tandem long-terminal-repeat circular unintegrated proviral (2-LTR) HIV-1 DNA, HIV-1 tat mRNA, and cell viability in the presence of three antiretroviral agents: recombinant soluble CD4 (rsCD4), zidovudine, and saquinavir. Interference with HIV-1 entry by rsCD4 decreased p24 antigen levels modestly, decreased HIV-1 gag by twofold, and 2-LTR was detectable at the end of the culture period. Inhibition of reverse transcription by zidovudine decreased p24 antigen levels modestly, decreased HIV-1 gag by 19-fold, and inhibited detection of 2-LTR HIV-1 DNA. The protease inhibitor, saquinavir, had the greatest overall effect, with the lowest levels of p24 antigen and HIV-1 gag, and inhibition of 2-LTR. There was no detection of tat mRNA in the saquinavir-treated cultures. In addition, cell viability was significantly higher in cultures treated with saquinavir. In these experiments, 2-LTR HIV-1 DNA was indicative of the relative inhibitory effects of three antiretroviral agents acting at different steps of the HIV-1 replication cycle. We demonstrated in vitro that 2-LTR HIV-1 DNA was a useful indicator of an antiretroviral drug in preventing new cell infection and could be utilized as a dynamic marker of drug efficacy in HIV-1-infected patients.

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Analysis of alternatively spliced human immunodeficiency virus type-1 mRNA species, one of which encodes a novel TAT-ENV fusion protein

Cited by 57 publications

References 62 publications

HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death

HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death

Infection of Macaca nemestrina brain with human immunodeficiency virus type 1

Unintegrated HIV-1 circular 2-LTR proviral DNA as a marker of recently infected cells: Relative effect of recombinant CD4, zidovudine, and saquinavir in vitro

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