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Quantitation of Simian Cytokine andβ-Chemokine mRNAs, using real-time reverse transcriptase-polymerase chain reaction: variations in expression during chronic primate lentivirus infection


Hofmann-Lehmann, Regina; Williams, Alison L; Swenerton, Ryan K; Li, Pei-Lin; Rasmussen, Robert A; Chenine, Agnès-Laurence; McClure, Harold M; Ruprecht, Ruth M (2002). Quantitation of Simian Cytokine andβ-Chemokine mRNAs, using real-time reverse transcriptase-polymerase chain reaction: variations in expression during chronic primate lentivirus infection. AIDS Research and Human Retroviruses, 18(9):627-639.

Abstract

Cytokines and β-chemokines are important mediators of the immune system and are expressed in many infectious diseases. To study cytokine and β-chemokine profiles during pathogenesis of lentiviral infection and progression to AIDS in rhesus macaques, we established new quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assays based on TaqMan chemistry. Using synthetic RNA standards, we quantified mRNAs of IL-2, IL-4, IL-6, IL-10, IL-12 p40, interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), RANTES, macrophage inflammatory protein 1α (MIP-1α), and MIP-1β in unstimulated peripheral blood mononuclear cells (PBMCs) and lymph nodes from macaques chronically infected with SIV or SHIV. Viremic monkeys with decreased CD4+ T cell counts (<500 cells/μl) had significantly higher IL-10 mRNA expression than uninfected controls, which parallels the findings in HIV-1-infected humans. In addition, MIP-1α, MIP-1β, and RANTES mRNA expression increased in viremic monkeys with decreased CD4+ T cell counts; gene expression was inversely correlated with CD4+ T cell counts, but not viral load. The newly established quantitative real-time RT-PCR assays will allow the determination of cytokine and β-chemokine patterns in rhesus macaques in studies of microbial pathogenesis or vaccine development.

Abstract

Cytokines and β-chemokines are important mediators of the immune system and are expressed in many infectious diseases. To study cytokine and β-chemokine profiles during pathogenesis of lentiviral infection and progression to AIDS in rhesus macaques, we established new quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assays based on TaqMan chemistry. Using synthetic RNA standards, we quantified mRNAs of IL-2, IL-4, IL-6, IL-10, IL-12 p40, interferon γ (IFN-γ), tumor necrosis factor α (TNF-α), RANTES, macrophage inflammatory protein 1α (MIP-1α), and MIP-1β in unstimulated peripheral blood mononuclear cells (PBMCs) and lymph nodes from macaques chronically infected with SIV or SHIV. Viremic monkeys with decreased CD4+ T cell counts (<500 cells/μl) had significantly higher IL-10 mRNA expression than uninfected controls, which parallels the findings in HIV-1-infected humans. In addition, MIP-1α, MIP-1β, and RANTES mRNA expression increased in viremic monkeys with decreased CD4+ T cell counts; gene expression was inversely correlated with CD4+ T cell counts, but not viral load. The newly established quantitative real-time RT-PCR assays will allow the determination of cytokine and β-chemokine patterns in rhesus macaques in studies of microbial pathogenesis or vaccine development.

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31 citations in Scopus®
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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinary Clinic > Department of Farm Animals
Dewey Decimal Classification:570 Life sciences; biology
630 Agriculture
Language:English
Date:2002
Deposited On:30 Nov 2016 14:07
Last Modified:08 Dec 2017 21:10
Publisher:Mary Ann Liebert
ISSN:0889-2229
Publisher DOI:https://doi.org/10.1089/088922202760019329
PubMed ID:12079558

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