Molecular characterization of woodchuck CD4 (wCD4) and production of a depletion monoclonal antibody against wCD4

• Published in: Molecular immunology 2013-11, Vol.56 (1-2), p.64-71
• Main Authors: Yang, Yinke, Zhang, Xiaoyong, Zhang, Chunyan, Tao, Yuanqing, Fan, Wei, Wang, Zhongdong, Wang, Hu, Lu, Mengji, Yang, Dongliang, Fiedler, Melanie, Wang, Baoju
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - metabolism; Antibodies, Monoclonal - pharmacology; Blotting, Western; CD4; CD4 Antigens - genetics; CD4 Antigens - immunology; CD4 Antigens - metabolism; CD4-Positive T-Lymphocytes - cytology; CD4-Positive T-Lymphocytes - immunology; CD4-Positive T-Lymphocytes - metabolism; Cell Proliferation - drug effects; Cloning, Molecular; Concanavalin A - immunology; Concanavalin A - pharmacology; Depletion antibody; DNA, Complementary - chemistry; DNA, Complementary - classification; DNA, Complementary - genetics; Electrophoresis, Polyacrylamide Gel; Hepatitis B virus; Marmota - genetics; Marmota - immunology; Marmota - metabolism; Marmota himalayana; Marmota monax; Molecular Sequence Data; Phylogeny; Primates; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Woodchuck
• ISSN: 0161-5890; 1872-9142
• DOI: 10.1016/j.molimm.2013.04.006

•The complete coding sequence of the woodchuck CD4 molecule was obtained and analyzed.•Two pAbs and four mAbs against wCD4 were produced and identified.•Anti-wCD4 mAb G2 effectively suppressed or depleted CD4 T cells in vitro and vivo.•This provides a basis for immune-related studies of HBV infection in woodchucks. 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.