Hidaka Y, Hayashi Y, Fisfalen M E, Suzuki S, Takeda T, Refetoff S, DeGroot L J
Thyroid Study Unit, University of Chicago, Illinois 60637, USA.
Thyroid. 1996 Feb;6(1):23-8. doi: 10.1089/thy.1996.6.23.
Thyroid peroxidase (TPO) is thought to be one of the pathological antigens in Hashimoto's thyroiditis. Epstein-Barr virus-transformed B cell lines (EBVL) can be used for antigen-presenting cells and target cells of cytotoxic T cells. To develop a model for endogenous TPO presentation in EBVL, we constructed a recombinant adenovirus carrying the TPO gene driven by the cytomegalovirus promoter (Ad5-TPO). Enzymatically active human TPO could be expressed in COS cells using Ad5-TPO. the peroxidase activity of the membrane extract from Ad5-TPO-infected COS cells was approximately 6 times higher than that from stably transfected TPO expressing CHO cells. TPO protein expression in the EBVL was analyzed by Western blotting technique. A band at approximately 110 kDa characteristic of hTPO was detected in EBVL infected with Ad5-TPO. hTPO expression in EBVL induced with adenovirus should facilitate understanding of T cell immunity to TPO in patients with autoimmune thyroid diseases.
甲状腺过氧化物酶（TPO）被认为是桥本甲状腺炎的病理抗原之一。爱泼斯坦-巴尔病毒转化的B细胞系（EBVL）可用于细胞毒性T细胞的抗原呈递细胞和靶细胞。为了建立EBVL中内源性TPO呈递的模型，我们构建了一种携带由巨细胞病毒启动子驱动的TPO基因的重组腺病毒（Ad5-TPO）。使用Ad5-TPO可在COS细胞中表达具有酶活性的人TPO。Ad5-TPO感染的COS细胞的膜提取物的过氧化物酶活性比稳定转染的表达TPO的CHO细胞高约6倍。通过蛋白质印迹技术分析EBVL中的TPO蛋白表达。在感染Ad5-TPO的EBVL中检测到一条约110 kDa的hTPO特征带。腺病毒诱导的EBVL中的hTPO表达应有助于理解自身免疫性甲状腺疾病患者对TPO的T细胞免疫。
