MOE Key Laboratory of Aquatic Product Safety/State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, People's Republic of China.
Dev Comp Immunol. 2012 Jan;36(1):39-50. doi: 10.1016/j.dci.2011.06.002. Epub 2011 Jun 26.
In vertebrates, the tumor necrosis factor (TNF)-receptor (TNFR) system participates in diverse physiological and pathological events, such as inflammation and protective immune responses to microbial infections. There are few reports about the role of the invertebrate TNF-TNFR system in immune responses. Here, we isolated and characterized the TNF superfamily (LvTNFSF) gene, TNFR superfamily (LvTNFRSF) gene and lipopolysaccharide-induced TNF-α factor (LvLITAF) gene from Litopenaeus vannamei. LvTNFSF consists of 472 amino acids with a conserved C-terminal TNF domain and has 89.8% identity with the Marsupenaeus japonicus TNF superfamily gene. LvTNFRSF consists of 296 amino acids with a conserved TNFR domain and has 18.0% identity with Chlamys farreri TNFR, 14.6% identity with Drosophila melanogaster Wengen and 14.6% identity with Homo sapiens TNFR1. LvLITAF consists of 124 amino acids with the LITAF domain and shows 62.6% identity with D. melanogaster LITAF and 32.3% identity with H. sapiens LITAF. The promoter region of LvTNFSF was cloned and used to construct a luciferase reporter. In Drosophila S2 cells, the promoter of LvTNFSF can be activated by LvLITAF, L. vannamei NF-κB family proteins (LvRelish and LvDorsal) and LvSTAT. Unlike its mammalian counterparts, LvTNFRSF could not activate the NF-κB pathway in Drosophila S2 cells. Using real-time quantitative PCR, we obtained expression profiles of LvTNFSF, LvTNFRSF and LvLITAF in the gill, intestine and hepatopancreas of L. vannamei after challenge with Gram-negative Vibrio alginolyticus, Gram-positive Staphylococcus aureus, the fungus Candida albicans and white spot syndrome virus (WSSV). Taken together, our results reveal that LvTNFSF, LvTNFRSF and LvLITAF may be involved in shrimp immune responses to pathogenic infections.
在脊椎动物中，肿瘤坏死因子（TNF）受体（TNFR）系统参与多种生理和病理事件，如炎症和对微生物感染的保护性免疫反应。关于无脊椎动物 TNF-TNFR 系统在免疫反应中的作用的报道很少。在这里，我们从凡纳滨对虾中分离并鉴定了 TNF 超家族（LvTNFSF）基因、TNFR 超家族（LvTNFRSF）基因和脂多糖诱导的 TNF-α因子（LvLITAF）基因。LvTNFSF 由 472 个氨基酸组成，具有保守的 C 末端 TNF 结构域，与日本沼虾 TNF 超家族基因的同源性为 89.8%。LvTNFRSF 由 296 个氨基酸组成，具有保守的 TNFR 结构域，与中国蛤蜊 TNFR 的同源性为 18.0%，与果蝇 Wengen 的同源性为 14.6%，与人类 TNFR1 的同源性为 14.6%。LvLITAF 由 124 个氨基酸组成，具有 LITAF 结构域，与果蝇 LITAF 的同源性为 62.6%，与人类 LITAF 的同源性为 32.3%。LvTNFSF 的启动子区域被克隆并用于构建荧光素酶报告基因。在果蝇 S2 细胞中，LvLITAF、凡纳滨对虾 NF-κB 家族蛋白（LvRelish 和 LvDorsal）和 LvSTAT 可以激活 LvTNFSF 的启动子。与哺乳动物不同的是，LvTNFRSF 不能激活果蝇 S2 细胞中的 NF-κB 途径。通过实时定量 PCR，我们获得了凡纳滨对虾鳃、肠和肝胰腺中 LvTNFSF、LvTNFRSF 和 LvLITAF 在革兰氏阴性弧菌、革兰氏阳性金黄色葡萄球菌、真菌白色念珠菌和白斑综合征病毒（WSSV）感染后的表达谱。综上所述，我们的结果表明 LvTNFSF、LvTNFRSF 和 LvLITAF 可能参与了对虾对致病性感染的免疫反应。
