Expression of thyroid peroxidase in EBV-transformed B cell lines using adenovirus.
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.
Hidaka
Y
Y
Thyroid Study Unit, University of Chicago, Illinois 60637, USA.
Hayashi
Y
Y
Fisfalen
M E
ME
Suzuki
S
S
Takeda
T
T
Refetoff
S
S
DeGroot
L J
LJ
D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology; D001402:B-Lymphocytes / Q000821:virology*; D015153:Blotting, Western; D002460:Cell Line; D002472:Cell Transformation, Viral; D004279:DNA, Viral / Q000096:biosynthesis; D004854:Herpesvirus 4, Human; D006801:Humans; D007453:Iodide Peroxidase / Q000096:biosynthesis* / Q000235:genetics; D008156:Luciferases / Q000032:analysis; D011401:Promoter Regions, Genetic / Q000235:genetics; D011995:Recombination, Genetic; D019065:Virus Assembly; D014779:Virus Replication
D004279:DNA, Viral; D007453:Iodide Peroxidase; D008156:Luciferases