The fibrous form of intracellular inclusion bodies in recombinant variant fibrinogen-producing cells is specific to the hepatic fibrinogen storage disease-inducible variant fibrinogen.
Fibrinogen storage disease (FSD) is a rare disorder that is characterized by the accumulation of fibrinogen in hepatocytes and induces liver injury. Six mutations in the γC domain (γG284R, γT314P, γD316N, the deletion of γG346-Q350, γG366S, and γR375W) have been identified for FSD. Our group previously established γ375W fibrinogen-producing Chinese hamster ovary (CHO) cells and observed aberrant large granular and fibrous forms of intracellular inclusion bodies. The aim of this study was to investigate whether fibrous intracellular inclusion bodies are specific to FSD-inducible variant fibrinogen. Thirteen expression vectors encoding the variant γ-chain were stably or transiently transfected into CHO cells expressing normal fibrinogen Aα- and Bβ-chains or HuH-7 cells, which were then immunofluorescently stained. Six CHO and HuH-7 cell lines that transiently produced FSD-inducible variant fibrinogen presented the fibrous (3.2-22.7 and 2.1-24.5%, respectively) and large granular (5.4-25.5 and 7.7-23.9%) forms of intracellular inclusion bodies. Seven CHO and HuH-7 cell lines that transiently produced FSD-non-inducible variant fibrinogen only exhibit the large granular form. These results demonstrate that transiently transfected variant fibrinogen-producing CHO cells and inclusion bodies of the fibrous form may be useful in non-invasive screening for FSD risk factors for FSD before its onset.
Department of Laboratory Medicine, Graduate School of Medicine, Shinshu University, Matsumoto, Japan.|Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Shinpei
S
Arai
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Naoko
N
Ogiwara
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Saki
S
Mukai
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Yuka
Y
Takezawa
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Mitsutoshi
M
Sugano
Department of Laboratory Medicine, Graduate School of Medicine, Shinshu University, Matsumoto, Japan.
Takayuki
T
Honda
Department of Health and Medical Sciences, Graduate School of Medicine, Shinshu University, Matsumoto, Japan. nobuoku@shinshu-u.ac.jp.|Laboratory of Clinical Chemistry and Immunology, Department of Biomedical Laboratory Sciences, School of Health Sciences, Shinshu University, 3-1-1 Asahi, Matsumoto, 390-8621, Japan. nobuoku@shinshu-u.ac.jp.
Nobuo
N
Okumura
D019943:Amino Acid Substitution; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005340:Fibrinogen / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology; D006801:Humans; D002479:Inclusion Bodies / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008107:Liver Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D020125:Mutation, Missense; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; D005340:Fibrinogen
Fibrinogen storage disease; Hypofibrinogenemia; Immunofluorescence; Intracellular inclusion bodies; Recombinant fibrinogen