Transcriptional profiling of TLR-4/7/8-stimulated guinea pig splenocytes and whole blood by bDNA assay

被引：15

作者：

Ching, Lance K. ^{[1
]}

Mompoint, Farah ^{[1
]}

Guderian, Jeffrey A. ^{[1
]}

Picone, Alex ^{[1
]}

Orme, Ian M. ^{[2
]}

Coler, Rhea N. ^{[1
,3
]}

Reed, Steven G. ^{[1
,3
,4
]}

Baldwin, Susan L. ^{[1
]}

机构：

[1] Infect Dis Res Inst, Seattle, WA 98104 USA

[2] Colorado State Univ, Coll Vet Med & Biomed Sci, Dept Microbiol Immunol & Pathol, Ft Collins, CO 80523 USA

[3] Univ Washington, Dept Global Hlth, Seattle, WA 98195 USA

[4] Immune Design Corp, Seattle, WA 98104 USA

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2011年 / 373卷 / 1-2期

基金：

美国国家卫生研究院;

关键词：

Toll-like receptors; Branched DNA; QuantiGene assay; Innate immunity; Cytokines; Guinea pig; DNA SIGNAL AMPLIFICATION; C VIRUS-RNA; HIV-1; RNA; PERFORMANCE-CHARACTERISTICS; TYPE-1; HCV RNA; QUANTIFICATION; EXPRESSION; PLASMA; QUANTITATION;

D O I：

10.1016/j.jim.2011.07.021

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Toll-like receptor (TLR) agonists are currently being examined as adjuvants for vaccines, with several lead candidates now in licensed products or in late-stage clinical development Guinea pigs are widely used for preclinical testing of drugs and vaccines; however, evaluation of TLR agonists in this model is hindered by the limited availability of immunological tools and reagents. In this study, we validated the use of a branched-chain DNA (bDNA) assay known as the QuantiGene Plex 2.0 Reagent System for measuring innate cytokine and chemokine mRNA levels following TLR stimulation of guinea pig cells. Gene expression for T-helper-1 (Th1) polarizing cytokines (TNF-alpha, IL-1 beta, IL-12) and chemokines (CXCL1, CCL2) was upregulated following ex vivo stimulation of guinea pig splenocytes and whole blood with TLR-4 or TLR-7/8 agonists. These data confirm the utility of the QuantiGene system both as an alternative to RT-PCR for measuring transcript levels and as a high-throughput screening tool for dissecting the immunological response to TLR stimulation in guinea pigs. Overall, the QuantiGene platform is reliable, reproducible, and sensitive. These agonists have the potential to be used as adjuvant components in vaccines against various pathogens. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：54 / 62

页数：9

共 26 条

[21] RAPID AND PRECISE QUANTIFICATION OF HIV-1 RNA IN PLASMA USING A BRANCHED DNA SIGNAL AMPLIFICATION ASSAY [J].