Huang San-Yuan, Chen Yu-Hui, Teng Shih-Hua, Chen I-Chung, Ho Lin-Lin, Tu Ching-Fu
Division of Biotechnology, Animal Technology Institute Taiwan, Miaoli, Taiwan.
Proteomics. 2006 Nov;6(21):5815-25. doi: 10.1002/pmic.200600054.
Matching donor and recipient human leucocyte antigen (HLA-II) could conquer cell-mediated rejection following transplantation. Transgenic pigs carrying HLA genes that "humanize" porcine organs, tissues, and cells were successfully generated. This study further clarifies the effect of HLA-DR transgenes on lymphocyte protein expression, via a proteomic approach. Lymphocytes were isolated from two HLA-DR transgenic pigs and three nontransgenic littermates on 157 d after birth. Soluble protein of 1x10(7) cells was separated using 2-DE. In total, 301 colloidal CBB-stained protein spots detected on all five 2-D gels were quantified. Thirty-three proteins were differentially expressed by a factor of 1.5. These proteins were subsequently identified by MALDI-TOF MS and MALDI-TOF/TOF MS/MS. These proteins were sorted into the following categories: chaperones, T-lymphocyte function, DNA/RNA processing, cytoskeleton-associated proteins, signal transduction, enzymes, and unknown. Previous studies have suggested that some of the identified proteins are associated with lymphocyte activation/proliferation. The identities of the unidentified spots and the systematic effect of these up- and down-regulated proteins on T-cell function in HLA-DR transgenic pigs require further exploration.
匹配供体和受体的人类白细胞抗原（HLA-II）可以克服移植后的细胞介导排斥反应。携带使猪器官、组织和细胞“人源化”的HLA基因的转基因猪已成功培育出来。本研究通过蛋白质组学方法进一步阐明了HLA-DR转基因对淋巴细胞蛋白质表达的影响。在出生后157天，从两头HLA-DR转基因猪和三只非转基因同窝仔猪中分离淋巴细胞。使用二维电泳分离1×10⁷个细胞的可溶性蛋白质。在所有五块二维凝胶上共检测到301个考马斯亮蓝染色的蛋白质斑点并进行定量分析。33种蛋白质的表达差异达到1.5倍。随后通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）和基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF/TOF MS/MS）鉴定这些蛋白质。这些蛋白质分为以下几类：伴侣蛋白、T淋巴细胞功能、DNA/RNA加工、细胞骨架相关蛋白、信号转导、酶和未知蛋白。先前的研究表明，一些已鉴定的蛋白质与淋巴细胞活化/增殖有关。在HLA-DR转基因猪中，未鉴定斑点的身份以及这些上调和下调蛋白质对T细胞功能的系统性影响需要进一步探索。
