Nakachi N, Matsunaga K, Klein T W, Friedman H, Yamamoto Y
Department of Medical Microbiology and Immunology, University of South Florida College of Medicine, Tampa, Florida 33612, USA.
Infect Immun. 2000 Oct;68(10):6069-72. doi: 10.1128/IAI.68.10.6069-6072.2000.
The cDNA expression array technique is a powerful tool to determine, at one time from many genes, specific gene messages modulated by infection. In the present study, we identified genes modulated in response to virulent versus avirulent Legionella pneumophila infection of the alveolar macrophage cell line MH-S by the cDNA expression array technique. Many macrophage genes were found to be modulated after 5 h of in vitro infection with L. pneumophila. In particular, it was found that the monocyte chemotactic protein 3 (MCP-3) gene expression was significantly induced by infection with virulent L. pneumophila but not with avirulent L. pneumophila. In contrast, other chemokine genes, such as macrophage inflammatory protein (MIP) 1alpha, were induced by both virulent and avirulent L. pneumophila. Reverse transcription (RT)-PCR assay of total RNA isolated from macrophages infected with the bacteria for 5 or 24 h confirmed the differential induction of the chemokine genes by virulent versus avirulent L. pneumophila. Thus, the cDNA expression array technique readily revealed differential induction by L. pneumophila infection of select chemokine genes of macrophages from more than 1,100 genes. These results also indicate that certain chemokine genes may be selectively induced by virulent bacteria.
cDNA表达阵列技术是一种强大的工具，可一次性从众多基因中确定受感染调节的特定基因信息。在本研究中，我们通过cDNA表达阵列技术，鉴定了肺泡巨噬细胞系MH-S在感染强毒与无毒嗜肺军团菌后被调节的基因。发现许多巨噬细胞基因在体外被嗜肺军团菌感染5小时后受到调节。特别地，发现单核细胞趋化蛋白3（MCP-3）基因表达在感染强毒嗜肺军团菌后显著诱导，而感染无毒嗜肺军团菌后未诱导。相反，其他趋化因子基因，如巨噬细胞炎性蛋白（MIP）1α，在强毒和无毒嗜肺军团菌感染后均被诱导。对感染细菌5或24小时的巨噬细胞分离的总RNA进行逆转录（RT）-PCR分析，证实了强毒与无毒嗜肺军团菌对趋化因子基因的差异诱导。因此，cDNA表达阵列技术很容易揭示嗜肺军团菌感染对来自1100多个基因中的巨噬细胞特定趋化因子基因的差异诱导。这些结果还表明，某些趋化因子基因可能被强毒细菌选择性诱导。
