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QCDCR配制的CpG ODN 2007对尼罗罗非鱼抗海豚链球菌的功效及上调基因的鉴定

Efficacy of QCDCR formulated CpG ODN 2007 in Nile tilapia against Streptococcus iniae and identification of upregulated genes.

作者信息

Pridgeon Julia W, Klesius Phillip H, Mu Xingjiang, Yancey Robert J, Kievit Michele S, Dominowski Paul J

机构信息

Aquatic Animal Health Research Unit, USDA-ARS, 990 Wire Road, Auburn, AL 36832, USA.

出版信息

Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):179-90. doi: 10.1016/j.vetimm.2011.11.001. Epub 2011 Nov 11.

DOI:10.1016/j.vetimm.2011.11.001
PMID:22129787
Abstract

The potential of using a QCDCR (quilA:cholesterol:dimethyl dioctadecyl ammonium bromide:carbopol:R1005 glycolipid) formulated CpG oligodeoxynucleotide (ODN), ODN 2007, to confer protection in Nile tilapia against Streptococcus iniae infection was evaluated in this study. At two days post treatment, QCDCR formulated ODN 2007 elicited significant (P<0.05) protection to Nile tilapia, with relative percent survival of 63% compared to fish treated by QCDCR alone. To understand the molecular mechanisms involved in the protective immunity elicited by ODN 2007, suppression subtractive cDNA hybridization technique was used to identify upregulated genes induced by ODN 2007. A total of 69 expressed sequence tags (ESTs) were identified from the subtractive cDNA library. Quantitative PCR revealed that 44 ESTs were significantly (P<0.05) upregulated by ODN 2007, including 29 highly (>10-fold) and 15 moderately (<10-fold) upregulated ESTs. Of all ESTs, putative peroxisomal sarcosine oxidase was upregulated the highest. The 69 ESTs only included six genes that had putative functions related to immunity, of which only two (putative glutaredoxin-1 and carboxypeptidase N catalytic chain) were confirmed to be significantly upregulated. Our results suggest that the protection elicited by ODN 2007 is mainly through innate immune responses directly or indirectly related to immunity.

摘要

本研究评估了使用由QCDCR(刺桐毒素:胆固醇:二甲基二十八烷基溴化铵:卡波姆:R1005糖脂)配制的CpG寡脱氧核苷酸(ODN)——ODN 2007,来保护尼罗罗非鱼免受海豚链球菌感染的潜力。在处理后两天,用QCDCR配制的ODN 2007对尼罗罗非鱼产生了显著(P<0.05)的保护作用,与仅用QCDCR处理的鱼相比,相对存活率为63%。为了解ODN 2007引发的保护性免疫所涉及的分子机制,采用抑制性消减cDNA杂交技术来鉴定由ODN 2007诱导上调的基因。从消减cDNA文库中总共鉴定出69个表达序列标签(EST)。定量PCR显示,44个EST被ODN 2007显著(P<0.05)上调,包括29个高度(>10倍)上调和15个中度(<10倍)上调的EST。在所有EST中,假定的过氧化物酶体肌氨酸氧化酶上调幅度最大。这69个EST中仅包括6个与免疫相关的具有假定功能的基因,其中只有2个(假定的谷氧还蛋白-1和羧肽酶N催化链)被证实显著上调。我们的结果表明,ODN 2007引发的保护作用主要是通过与免疫直接或间接相关的先天免疫反应实现的。

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