Peatman Eric, Baoprasertkul Puttharat, Terhune Jeffery, Xu Peng, Nandi Samiran, Kucuktas Huseyin, Li Ping, Wang Shaolin, Somridhivej Benjaporn, Dunham Rex, Liu Zhanjiang
The Fish Molecular Genetics and Biotechnology Laboratory, Department of Fisheries and Allied Aquacultures and Program of Cell and Molecular Biosciences, Aquatic Genomics Unit, Auburn University, 203 Swingle Hall, Auburn, AL 36849, USA.
Dev Comp Immunol. 2007;31(11):1183-96. doi: 10.1016/j.dci.2007.03.003. Epub 2007 Apr 9.
The acute phase response (APR) is a set of metabolic and physiological reactions occurring in the host in response to tissue infection or injury and is a crucial component of the larger innate immune response. The APR is best characterized by dramatic changes in the concentration of a group of plasma proteins known as acute phase proteins (APPs) which are synthesized in the liver and function in a wide range of immunity-related activities. Utilizing a new high-density in situ oligonucleotide microarray, we have evaluated the APR in channel catfish liver following infection with Edwardsiella ictaluri, a bacterial pathogen that causes enteric septicemia of catfish. Our catfish microarray design (28K) builds upon a previous 19K channel catfish array by adding recently sequenced immune transcripts from channel catfish along with 7159 unique sequences from closely related blue catfish. The analysis of microarray results using a traditional 2-fold change in gene expression cutoff and a 10% false-discovery rate revealed a well-developed APR in catfish, with particularly high upregulation (>50-fold) of genes involved in iron homeostasis (i.e. intelectin, hemopexin, haptoglobin, ferritin, and transferrin). Other classical APP genes upregulated greater than 2-fold included coagulation factors, proteinase inhibitors, transport proteins, and complement components. Upregulation of the majority of the complement cascade was observed including the membrane attack complex components and complement inhibitors. A number of pathogen recognition receptors (PRRs) and chemokines were also differentially expressed in the liver following infection. Independent testing of a selection of differentially expressed genes with real-time RT-PCR confirmed microarray results.
急性期反应（APR）是宿主针对组织感染或损伤而发生的一系列代谢和生理反应，是更大的先天免疫反应的关键组成部分。APR的最佳特征是一组称为急性期蛋白（APPs）的血浆蛋白浓度发生显著变化，这些蛋白在肝脏中合成，并在广泛的免疫相关活动中发挥作用。利用一种新的高密度原位寡核苷酸微阵列，我们评估了斑点叉尾鮰肝脏在感染鮰爱德华氏菌后的急性期反应，鮰爱德华氏菌是一种导致斑点叉尾鮰肠道败血症的细菌病原体。我们的斑点叉尾鮰微阵列设计（28K）是在先前的19K斑点叉尾鮰阵列的基础上构建的，增加了最近测序的斑点叉尾鮰免疫转录本以及来自亲缘关系密切的蓝鲶的7159个独特序列。使用传统的基因表达截止值2倍变化和10%的错误发现率对微阵列结果进行分析，发现斑点叉尾鮰有发达的急性期反应，参与铁稳态的基因（即凝集素、血红素结合蛋白、触珠蛋白、铁蛋白和转铁蛋白）有特别高的上调（>50倍）。上调超过2倍的其他经典APP基因包括凝血因子、蛋白酶抑制剂、转运蛋白和补体成分。观察到补体级联反应的大部分上调，包括膜攻击复合物成分和补体抑制剂。感染后，肝脏中一些病原体识别受体（PRRs）和趋化因子也有差异表达。用实时RT-PCR对选定的差异表达基因进行独立测试，证实了微阵列结果。
