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Expression of thyroid peroxidase in EBV-transformed B cell lines using adenovirus
Y Hidaka1, Y Hayashi, M E Fisfalen
1Thyroid Study Unit, University of Chicago, Illinois 60637, USA.
Abstract:
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.