Molecular cloning, characterization and expression analysis of Tim-3 and Galectin-9 in the woodchuck model.
In recent years, a critical role for T cell immunoglobulin mucin domain 3 (Tim-3) and its ligand Galectin-9 (Gal-9) has emerged in infectious disease, autoimmunity and cancer. Manipulating this immune checkpoint may have immunotherapeutic potential and could represent an alternative approach for improving immune responses to viral infections and cancer. The woodchuck (Marmot monax) infected by woodchuck hepatitis virus (WHV) represents an informative animal model to study HBV infection and HCC. In the current study, the cDNA sequences of woodchuck Tim-3 and Gal-9 were cloned, sequenced and characterized. The extracellular domain of Tim-3 cDNA sequence consisted of 576bp coding sequence (CDS) that encoded 192 amino acids. The 1076bp full-length Gal-9 cDNA sequence consisted of 1059bp coding sequence (CDS) that encoded 352 amino acids with a molecular weight of 39.7kDa. The phylogenetic tree analysis revealed that the woodchuck Tim-3 and Gal-9 had the closest genetic relationship with Ictidomys tridecemlineatus. The result of quantification PCR analysis showed that ubiquitous expression of Gal-9 but not Tim-3 in different tissues of naive woodchucks. Elevated liver Gal-9 expression was observed in woodchucks with chronic WHV infection. Moreover, a polyclonal antibody against the extracellular domain of woodchuck Tim-3 were generated and identified by flow cytometry. Our results serve as a foundation for further insight into the role of Tim-3/Galectin-9 signaling pathway in viral hepatitis and HCC in the woodchuck model.
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Yanan
Y
Liu
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Junzhong
J
Wang
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Lu
L
Wang
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Baoju
B
Wang
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Shangqing
S
Yang
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Qin
Q
Wang
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Jinzhuo
J
Luo
Institute of Infection and Immunology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Xuemei
X
Feng
Institute of Infection and Immunology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Xuecheng
X
Yang
Institute of Infection and Immunology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Yinping
Y
Lu
Institute for Virology, University Hospital of Essen, University of Duisburg-Essen, Germany.
Michael
M
Roggendorf
Institute for Virology, University Hospital of Essen, University of Duisburg-Essen, Germany.
Mengji
M
Lu
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Institute of Infection and Immunology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Electronic address: dlyang@hust.edu.cn.
Dongliang
D
Yang
Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Institute of Infection and Immunology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. Electronic address: jialiu77@hotmail.com.
Jia
J
Liu
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D037161:Galectins / Q000096:biosynthesis / Q000235:genetics*; D018347:Hepadnaviridae Infections / Q000235:genetics / Q000276:immunology; D000072597:Hepatitis A Virus Cellular Receptor 2 / Q000096:biosynthesis / Q000235:genetics*; D017919:Hepatitis B Virus, Woodchuck; D006524:Hepatitis, Viral, Animal / Q000235:genetics / Q000276:immunology; D008392:Marmota / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D059467:Transcriptome
D018076:DNA, Complementary; D037161:Galectins; D000072597:Hepatitis A Virus Cellular Receptor 2
Expression; Galectin-9; Hepatitis; Sequence analysis; Tim-3; Woodchuck