title: Development of a Cell Marker ELISA for the Detection of Goose T Cell Surface CD8α Molecules.
abstract: CD8 molecule is a key marker on T cell surface and is connected with the antigen recognition and activation of T lymphocytes. In order to provide a detection method for quantifying goose CD8α expression, this study raised the protein and antibody for goose CD8α and developed a feasible cell marker enzyme-linked immunoabsorbent assay (ELISA) method. Recombinant protein of the extracellular region gene of goCD8α was expressed in prokaryotic expression system, and specific polyclonal antibodies for goCD8α were raised and purified, which was further confirmed by Western-blot, immunofluorescence assay (IFA), and immunohistochemistry (IHC). A cell marker ELISA was established and optimized to detect the change of goCD8α expression between goose parvovirus (GPV)-infected and mock-infected goose peripheral blood mononuclear cells (PBMCs), which is consistent with our previously results of real-time quantitative PCR (qPCR). Cell marker ELISA can provide a new method to detect goCD8α in protein level and in a sensitive, specific, and simple way. This may provide a convenient and novel method for the detection of goCD8α expression.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.
authors.forename: Beibei
authors.initials: B
authors.lastname: Cheng
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China. sophia_cs@163.com.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. sophia_cs@163.com.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. sophia_cs@163.com.
authors.forename: Shun
authors.initials: S
authors.lastname: Chen
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Mingshu
authors.initials: M
authors.lastname: Wang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Renyong
authors.initials: R
authors.lastname: Jia
authors.affiliation: Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Dekang
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.
authors.forename: Mafeng
authors.initials: M
authors.lastname: Liu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Kunfeng
authors.initials: K
authors.lastname: Sun
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Chen
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, No. 211 Huimin Road, Wenjiang District, Chengdu City, Sichuan Province, 611130, China. chenganchun@vip.163.com.|Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. chenganchun@vip.163.com.|Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. chenganchun@vip.163.com.
authors.forename: Anchun
authors.initials: A
authors.lastname: Cheng
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000954:Antigens, Surface / Q000235:genetics / Q000302:isolation & purification*; D016827:CD8 Antigens / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005777:Geese / Q000235:genetics / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D017992:Parvovirus / Q000276:immunology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013601:T-Lymphocytes / Q000276:immunology; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000914:Antibodies, Viral; D000954:Antigens, Surface; D016827:CD8 Antigens; C431698:CD8alpha antigen; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7
keywords: Cell marker ELISA; Goose CD8α; Polyclonal antibody; Prokaryotic expression