Ⅱ型成对免疫球蛋白样受体(Paired immunoglobulin-like type 2 receptors, PILRs)是免疫球蛋白超家族成员之一,包括α和β两个亚型。PILRα在机体抵抗病原体入侵的免疫反应中发挥着重要作用,但目前尚未见关于猪PILRα的报道。为了分析其在猪抗病育种中的作用,本文克隆了猪PILRα编码基因(PILRA)并鉴定变异剪接体,利用实时荧光定量PCR方法构建其组织表达谱和诱导表达谱。结果表明,成功克隆了猪PILRA基因的3个变异剪接体V1~V3(GenBank登录号：KJ143679~81),预测的蛋白质多肽链分别长271 aa、254 aa和283 aa,且都具有免疫球蛋白结构域。各变异剪接体均在脾脏中表达量最高,肝脏、肺脏次之,在心脏、肾脏、胃、肌肉、淋巴、大肠、小肠和膀胱等组织中表达量较低或检测不到。Poly(I:C)能显著诱导变异剪接体V1的表达,但几乎不影响V2、V3的表达。研究结果为进一步揭示PILRA在猪抗病育种中的作用提供了基础。
Paired immunoglobulin-like type 2 receptors (PILRs) are members of the immunoglobulin superfamily and composed of two subtypes, α and β. PILRα plays an important role in the immune response against invading pathogens, but so far there is no report on porcine PILRα. In order to analyze the potential role of PILRα in porcine disease-resistant breeding, we first cloned the PILRA gene (V1-V3, GenBank accession Nos. KJ143679-81) into pigs, and identified its three splice variants. Each variant conceptually translates into proteins of 271 amino acids (aa), 254aa and 283aa, respectively. Furthermore, quantitative real-time PCR was used to construct expression profiles of each variant in tissues and that induced by Poly(I:C). All three variants had the highest expression levels in the spleen, followed by liver and lung tissues. While levels were low or undetectable in the heart, kidney, stomach, muscle, lymph, large intestine, small intestine and bladder. Poly(I:C) significantly induced the expression of splice variant 1 (V1) of porcine PILRA, but hardly affected the expression of V2 and V3. The results lay a foundation for further study on the role of PILRA in porcine breeding and disease resistance.
[1] Takai T. Paired immunoglobulin-like receptors and their MHC class I recognition. Immunology , 2005, 115(4): 433-440.
[2] Wilson MD, Cheung J, Martindale DW, Scherer SW, Koop BF. Comparative analysis of the paired mmunoglobulin-like receptor ( PILR ) locus in six mammalian genomes: duplication, conversion, and the birth of new genes. Physiol Genomics , 2006, 27(3): 201-218.
[3] Lun YZ, Chi Q, Wang XL, Wang F, Sui W. Identification of paired immunoglobulin-like type 2 receptor α as hepatitis B virus DNA polymerase transactivated protein 1 interacting proteins. Mol Med Rep , 2014, 9(2): 720-724.
[4] Satoh T, Arase H. HSV-1 infection through inhibitory receptor, PILRalpha. Uirusu , 2008, 58(1): 27-36.
[5] Wang J, Fan Q, Satoh T, Arii J, Lanier LL, Spear PG, Kawaguchi Y, Arase H. Binding of herpes simplex virus glycoprotein B (gB) to paired immunoglobulin-like type 2 receptor α depends on specific sialylated O-linked glycans on gB. J Virol , 2009, 83(24): 13042-13045.
[6] Banerjee A, Stevenaert F, Pande K, Haghjoo E, Antonenko S, Gorman DM, Sathe M, McClanahan TK, Pierce R, Turner SP, Bigler ME, Phillips JH, Heyworth PG. Modulation of paired immunoglobulin-like type 2 receptor signaling alters the host response to Staphylococcus aureus-induced pneumonia. Infect Immun , 2010, 78(3): 1353-1363.
[7] Weber F, Wagner V, Rasmussen SB, Hartmann R, Paludan SR. Double-stranded RNA is produced by positive-strand RNA viruses and DNA viruses but not in detectable amounts by negative-strand RNA viruses. J Virol , 2006, 80(10): 5059-5064.
[8] 王秋实, 武春艳, 李朋, 邵思宇, 张姣, 朱蒙, 杨秀芹. 猪 GPR 84和 MPEG 1基因的分子克隆和表达分析. 畜牧兽医学报, 2014, 45(5): 699-705.
[9] Tato CM, Joyce-Shaikh B, Banerjee A, Chen Y, Sathe M, Ewald SE, Liu MR, Gorman D, McClanahan TK, Phillips JH, Heyworth PG, Cua DJ. The myeloid receptor PILRβ mediates the balance of inflammatory responses through regulation of IL-27 production. PLoS One , 2012, 7(3): e31680.
[10] Sun YL, Senger K, Baginski TK, Mazloom A, Chinn Y, Pantua H, Hamidzadeh K, Ramani SR, Luis E, Tom I, Sebrell A, Quinones G, Ma Y, Mukhyala K, Sai T, Ding JB, Haley B, Shadnia H, Kapadia SB, Gonzalez LC, Hass PE, Zarrin AA. Evolutionarily conserved paired immunoglobulin-like receptor α (PILRα) domain mediates its interaction with diverse sialylated ligands. J Biol Chem , 2012, 287(19): 15837-15850.
[11] Pan Q, Shai O, Lee LJ, Frey BJ, Blencowe BJ. Deep surveying of alternative splicing complexity in the human transcriptome by high-throughput sequencing. Nat Genet , 2008, 40(12): 1413-1415.
[12] Wang ET, Sandberg R, Luo SJ, Khrebtukova I, Zhang L, Mayr C, Kingsmore SF, Schroth GP, Burge CB. Alternative isoform regulation in human tissue transcriptomes. Nature , 2008, 456(7221): 470-476.
[13] Shiratori I, Ogasawara K, Saito T, Lanier LL, Arase H. Activation of natural killer cells and dendritic cells upon recognition of a novel CD99-like ligand by paired immunoglobulin-like type 2 receptor. J Exp Med , 2004, 199(4): 525-533.
