title: Molecular characterization of woodchuck CD4 (wCD4) and production of a depletion monoclonal antibody against wCD4.
abstract: CD4 T cells play an important role in the immune response against hepatitis B virus (HBV) infection. Woodchucks represent an excellent animal model to study HBV infection. In this study, we characterized the cDNA sequence of woodchuck CD4 (wCD4). The deduced wCD4 protein has four extracellular immunoglobulin-like domains comparable to the other mammalian CD4 molecules. The important extracellular cysteine residues and the intracellular tyrosine protein kinase-binding site of wCD4 are also conserved. The deduced wCD4 protein shows 53-63% identity with the counterparts of other mammalians. Phylogenetic analysis indicates that wCD4 is closely related with the counterparts of primates. Two polyclonal antibodies (pAbs) and four monoclonal Abs (mAbs) against wCD4 were produced. Two pAbs and one mAbs (G2) were found to effectively suppress ConA induced proliferation in vitro. Anti-wCD4 mAb G2 depleted 60% of CD4 cells from healthy woodchucks, while the remaining CD4 cells responded well to ConA stimulation. This work provides a basis for studying CD4 T cell mediated immune responses against HBV infection in the woodchuck model.
authors.affiliation: Department of Infectious Diseases, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Yinke
authors.initials: Y
authors.lastname: Yang
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Zhang
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhang
authors.forename: Yuanqing
authors.initials: Y
authors.lastname: Tao
authors.forename: Wei
authors.initials: W
authors.lastname: Fan
authors.forename: Zhongdong
authors.initials: Z
authors.lastname: Wang
authors.forename: Hu
authors.initials: H
authors.lastname: Wang
authors.forename: Mengji
authors.initials: M
authors.lastname: Lu
authors.forename: Dongliang
authors.initials: D
authors.lastname: Yang
authors.forename: Melanie
authors.initials: M
authors.lastname: Fiedler
authors.forename: Baoju
authors.initials: B
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D015153:Blotting, Western; D015704:CD4 Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000276:immunology / Q000494:pharmacology; D018076:DNA, Complementary / Q000737:chemistry / Q000145:classification / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008392:Marmota / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D018076:DNA, Complementary; D003208:Concanavalin A