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Identification of Differentially Expressed Genes in a Monkey Model of Allergic Asthma by Microarray Technology* FREE TO VIEW

Jun Zou, PhD; Simon Young, PhD; Feng Zhu, MD; Lu Xia, MS; Susan Skeans, BS; Yuntao Wan, MS; Luquan Wang, PhD; Terri McClanahan, PhD; Ferdous Gheyas, PhD; Din Wei, PhD; Charles Garlisi, PhD; James Jakway, PhD; Shelby Umland, PhD
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*From the Departments of Allergy (Drs. Zou, Young, Zhu, Garlisi, Jakway, and Umland, and Mss. Xia, Skeans, and Wan), Bioinformatics (Drs. Wang and Wei), and Biostatistics (Dr. Gheyas), Schering-Plough Research Institute, Kenilworth, NJ; and DNAX Research Institute (Dr. McClanahan), Palo Alto, CA.

Correspondence to: Jun Zou, PhD, Schering-Plough Research Institute, 2000 Galloping Hill Rd, Kenilworth, NJ 07033-0530; e-mail: jun.zou@spcorp.com

Chest. 2002;121(3_suppl):26S-27S. doi:10.1378/chest.121.3_suppl.26S
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Inhalation of the antigen Ascaris suum by allergic monkeys causes an immediate and delayed allergic reaction, including an influx of inflammatory cells into the lungs. The resulting bronchoconstriction and airway inflammation with eosinophilia are symptoms that are typical cases of human asthma. In order to identify the genes that play crucial roles in this disease process, we conducted a genome-wide gene expression profiling of allergic monkey lungs by using microarrays technology (Synteni; Fremont, CA). Individual monkeys were challenged by the inhalation of the A suum antigen or interleukin-4, and lung tissue was collected at 4, 18, or 24 h after the A suum challenge or 24 h after the interleukin-4 challenge. The results of the lung probes of each challenged monkey were compared with those from a probe prepared from a pool of control, unchallenged monkey lungs. Cluster analysis was used to study the data. More than 100 genes were found to be up-regulated or down-regulated by > 2.5-fold in at least one of the probe pairs. Among those > 100 genes, one quarter of them are novel genes and three quarters of them are known genes. One third of these known genes have been reported in the literature to be associated with human allergy or asthma. To validate the microarray results, the differential expression of a subset of the known genes was measured by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in the tissue used for probing the microarray as well as in the tissue of three additional monkeys (4 h post-A suum challenge). The confirmation of the differential expression in the original sample was found in most of the 18 genes tested by RT-PCR. In general, larger differences in expression levels were observed by RT-PCR than by microarray. Nine of these 18 genes showed a statistically significant difference when examined in multiple monkey lungs 4 h after the challenge (n = 4) compared to control lungs (n = 6). In conclusion, a set of genes differentially regulated and identified by microarray were confirmed by quantitative PCR, thus validating this approach. In addition, this study also demonstrated that the regulated expression of some genes associated with human asthma/allergy were also similarly regulated in the challenged lungs of allergic monkey.

Abbreviation: RT-PCR = reverse transcriptase-polymerase chain reaction




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