Application OF Branched DNA Signal Amplification to Monitor Human Immunodeficiency Virus Type 1 Burden in Human Plasma

Dewar, Robin; Highbarger, Helene; Sarmiento, Marion; Todd, John; Vasudevachari, M. B.; Davey, Richard T.; Kovacs, Joseph A.; Salzman, Norman P.; Lane, H. Clifford; Urdea, Mickey S.

doi:10.1093/infdis/170.5.1172

Cited by 169 publications

(78 citation statements)

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“…Total RNA was reverse transcribed into cDNA using Superscript III (Invitrogen, Carlsbad, CA), and the expression levels of CYP3A4, ABCB1, ABCC2, or ABCG2 or the endogenous control, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), were measured using 6-carboxyfluorescein (FAM)-labeled TaqMan gene expression assays and the primer-limited VIC/MGB probe for GAPDH (Life Technologies Inc., Burlington, Ontario, Canada) in the Mastercycler ep realplex 2S thermal cycler (Eppendorf Canada, Mississauga, Ontario, Canada). HIV RNA levels in intestinal tissue and plasma samples were measured by real-time qPCR as described previously (31,32).…”

Section: Methodsmentioning

confidence: 99%

HIV-1 Alters Intestinal Expression of Drug Transporters and Metabolic Enzymes: Implications for Antiretroviral Drug Disposition

Kis

Sankaran-Walters

Hoque

et al. 2016

Antimicrob Agents Chemother

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؉ ART-naive group than in uninfected subjects. qPCR analysis confirmed significantly lower expression of ABCC2/MRP2 in ART-naive subjects than in the control group, while CYP3A4 and ABCG2/BCRP showed a trend toward decreased expression. Protein expression of MRP2 and BCRP was also significantly lower in the HIV ؉ naive group than in the control group and was partially restored to baseline levels in HIV ؉ subjects receiving ART. In contrast, gene and protein expression of ABCB1/Pgp was significantly increased in HIV ؉ subjects on ART relative to HIV ؉ ART-naive subjects. These data demonstrate that the expression of drug-metabolizing enzymes and efflux transporters is significantly altered in therapy-naive HIV ؉ subjects and in those receiving ART. Since CYP3A4, Pgp, MRPs, and BCRP metabolize or transport many antiretroviral drugs, their altered expression with HIV infection may negatively impact drug pharmacokinetics in HIV ؉ subjects. This has clinical implications when using data from healthy volunteers to guide ART.

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Section: Methodsmentioning

confidence: 99%

HIV-1 Alters Intestinal Expression of Drug Transporters and Metabolic Enzymes: Implications for Antiretroviral Drug Disposition

Kis

Sankaran-Walters

Hoque

et al. 2016

Antimicrob Agents Chemother

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show abstract

“…Plasma HIV-1 RNA was determined by quantitative branched-chain DNA (bDNA; Chiron, Emeryville, CA) and ultrasensitive reverse transcriptase-PCR (RT-PCR; Roche Molecular Systems, Branchburg, NJ) assays. Levels were expressed as copies/ml, and the lower levels of sensitivity were 500 copies/ml (bDNA assay) and 50 copies/ml (RT-PCR assay) (12). Absolute blood CD4 ϩ T-cell counts were measured by consensus flow cytometry methodology.…”

Section: Methodsmentioning

confidence: 99%

Enhanced Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Nef-Specific T Cells Recognizing Multiple Variants in Early HIV-1 Infection

Malhotra¹,

Nolin³

et al. 2007

J Virol

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A human immunodeficiency virus (HIV)-preventive vaccine will likely need to induce broad immunity that can recognize antigens expressed within circulating strains. To understand the potentially relevant responses that T-cell based vaccines should elicit, we examined the ability of T cells from early infected persons to recognize a broad spectrum of potential T-cell epitopes (PTE) expressed by the products encoded by the HIV type 1 (HIV-1) nef gene, which is commonly included in candidate vaccines. T cells were evaluated for gamma interferon (IFN-␥) secretion using two peptide panels: subtype B consensus (CON) peptides and a novel peptide panel providing 70% coverage of PTE in subtype B HIV-1 Nef. Eighteen of 23 subjects' T cells recognized HIV-1 Nef. In one subject, Nef-specific T cells were detected with the PTE but not with the CON peptides. The greatest frequency of responses spanned Nef amino acids 65 to 103 and 113 to 147, with multiple epitope variants being recognized. Detection of both the epitope domain number and the response magnitude was enhanced using the PTE peptides. On average, we detected 2.7 epitope domains with the PTE peptides versus 1.7 domains with the CON peptides (P ‫؍‬ 0.0034). The average response magnitude was 2,169 spot-forming cells (SFC)/10 6 peripheral blood mononuclear cells (PBMC) with the PTE peptides versus 1,010 SFC/10 6 PBMC with CON peptides (P ‫؍‬ 0.0046). During early HIV-1 infection, Nef-specific T cells capable of recognizing multiple variants are commonly induced, and these responses are readily detected with the PTE peptide panel. Our findings suggest that Nef responses induced by a given vaccine strain before HIV-1 exposure may be sufficiently broad to recognize most variants within subtype B HIV-1.

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“…Plasma HIV-1 RNA was determined by quantitative branched-DNA assay (bDNA; Chiron Corp., Emeryville, California, USA) and ultrasensitive RT-PCR assay (Roche Molecular Systems, Branchburg, New Jersey, USA). Levels were expressed as copies per milliliter, and the lower levels of sensitivity were 500 copies/ml (bDNA assay) and 50 copies/ml (RT-PCR assay) (16). We report viral levels measured by the bDNA assay when there are more than 500 copies/ml and by the ultrasensitive RT-PCR assay when the bDNA assay was less than or equal to 500 copies/ml.…”

Section: Methodsmentioning

confidence: 99%

Role for HLA class II molecules in HIV-1 suppression and cellular immunity following antiretroviral treatment

Malhotra¹,

Holte²,

Dutta³

et al. 2001

J. Clin. Invest.

113

110

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Application OF Branched DNA Signal Amplification to Monitor Human Immunodeficiency Virus Type 1 Burden in Human Plasma

Cited by 169 publications

References 0 publications

HIV-1 Alters Intestinal Expression of Drug Transporters and Metabolic Enzymes: Implications for Antiretroviral Drug Disposition

HIV-1 Alters Intestinal Expression of Drug Transporters and Metabolic Enzymes: Implications for Antiretroviral Drug Disposition

Enhanced Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Nef-Specific T Cells Recognizing Multiple Variants in Early HIV-1 Infection

Role for HLA class II molecules in HIV-1 suppression and cellular immunity following antiretroviral treatment

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