Zhang Ran, Liu Rui, Wang Weilin, Xin Lusheng, Wang Lingling, Li Chenghua, Song Linsheng
School of Marine Sciences, Ningbo University, Ningbo, 315211, China.
Key laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.
Fish Shellfish Immunol. 2015 Jun;44(2):547-54. doi: 10.1016/j.fsi.2015.03.015. Epub 2015 Mar 23.
Interferons (IFNs) belong to class II helical cytokines family with pleiotropic biological activities, which have been demonstrated to play crucial roles in innate and adaptive immunity in vertebrates. In the present study, a novel IFN-like protein (designed CgIFNLP) was identified from oyster Crassostrea gigas, which contained an interferon domain from 14 to 97 amino acids showing low sequence similarities with vertebrates IFNs, but shared a similar three-dimensional structure with class II helical cytokines. The mRNA transcripts of CgIFNLP was detected in all the tested tissues including gonad, adductor muscle, hemocytes, mantle, gills, and hepatopancreas, with the highest expression level in gills (39-fold, P < 0.05). Moreover, the expression level of CgIFNLP mRNA in hemocytes increased significantly at 12 h (8.35-fold, P < 0.01) and 24 h (4.95-fold, P < 0.01) after poly (I: C) stimulation. After the treatments by recombinant CgIFNLP protein (rCgIFNLP) at different concentrations, the apoptosis and phagocytosis rates of oyster hemocytes increased obviously. The proliferation rate of L929 did not change obviously after incubation with rCgIFNLP for 72 h, but the proliferation rate of A549 abated significantly at 36 h and 48 h after incubation with rCgIFNLP. The results collectively suggested that the IFN-like molecule existed in oyster and it tended to present conserved functions rather than conserved amino acid sequence in comparison with vertebrate IFNs.
干扰素(IFNs)属于具有多效生物活性的II类螺旋细胞因子家族,已证明其在脊椎动物的先天免疫和适应性免疫中发挥关键作用。在本研究中,从太平洋牡蛎(Crassostrea gigas)中鉴定出一种新型干扰素样蛋白(命名为CgIFNLP),其含有一个14至97个氨基酸的干扰素结构域,与脊椎动物干扰素的序列相似性较低,但与II类螺旋细胞因子具有相似的三维结构。在所有测试组织中均检测到CgIFNLP的mRNA转录本,包括性腺、闭壳肌、血细胞、外套膜、鳃和肝胰腺,其中鳃中的表达水平最高(39倍,P < 0.05)。此外,在聚肌胞苷酸(poly (I: C))刺激后12小时(8.35倍,P < 0.01)和24小时(4.95倍,P < 0.01),血细胞中CgIFNLP mRNA的表达水平显著增加。用不同浓度的重组CgIFNLP蛋白(rCgIFNLP)处理后,牡蛎血细胞的凋亡率和吞噬率明显增加。与rCgIFNLP孵育72小时后,L929的增殖率没有明显变化,但与rCgIFNLP孵育36小时和48小时后,A549的增殖率显著降低。这些结果共同表明,牡蛎中存在干扰素样分子,与脊椎动物干扰素相比,它倾向于呈现保守的功能而非保守的氨基酸序列。