title: C3-Luc Cells Are an Excellent Model for Evaluation of Cellular Immunity following HPV16L1 Vaccination.
abstract: C3 and TC-1 are the two model cell lines most commonly used in studies of vaccines and drugs against human papillomavirus (HPV) infection. Because C3 cells contain both the HPV16 E and L genes, but TC-1 cells contain only the HPV16 E genes, C3 cells are usually used as the model cell line in studies targeting the HPV16 L protein. However, expression of the L1 protein is difficult to detect in C3 cells using common methods. In our study, Short tandem repeat analysis (STR) was used to demonstrate that C3 cells are indeed derived from mice, PCR results show that HPV16 L1, E6 and E7 genes were detected in C3 genomic DNA, and RT-PCR results demonstrated that L1 transcription had occurred in C3 cells. However, the expression of C3 protein was not found in the results of western blot and immunohistochemistry (IHC). Growth and proliferation of C3 were inhibited by mice spleen lymphocytes that had been immunized with a vaccine against HPV16L1. The luciferase gene was integrated into C3 cells, and it was confirmed that addition of the exogenous gene had no effect on C3 cells by comparing cell growth and tumor formation with untransformed cells. Cells stably expressing luciferase (C3-luc) were screened and subcutaneously injected into the mice. Tumors became established and were observed using a Spectrum Pre-clinical in Vivo Imaging System. Tumor size of mice in the different groups at various time points was calculated by counting photons. The sensitivity of the animals to the vaccine was quantified by statistical comparison. Ten or 30 days following injection of the C3-luc cells, tumor size differed significantly between the PBS and vaccine groups, indicating that C3 cells were susceptible to vaccination even after tumors were formed in vivo.
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.|National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, State Key Laboratory for Infectious Disease Prevention and Control, Beijing, China.
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Li
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.
authors.forename: He-Rong
authors.initials: HR
authors.lastname: Wang
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.
authors.forename: Zhi-Yi
authors.initials: ZY
authors.lastname: Zhou
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Luo
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Wang
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.
authors.forename: Xiang-Qian
authors.initials: XQ
authors.lastname: Xiao
authors.affiliation: Beijing Key Laboratory of Environmental and Viral Oncology, College of Life Science and Bio-Engineering, Beijing University of Technology, Beijing, China.
authors.forename: Yu-Bai
authors.initials: YB
authors.lastname: Zhou
authors.affiliation: National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, State Key Laboratory for Infectious Disease Prevention and Control, Beijing, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D000818:Animals; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology*; D063646:Carcinogenesis / Q000187:drug effects / Q000473:pathology; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D005814:Genes, Viral; D006801:Humans; D007111:Immunity, Cellular; D007150:Immunohistochemistry; D008156:Luciferases / Q000378:metabolism* / Q000494:pharmacology; D008810:Mice, Inbred C57BL; D018895:Microsatellite Repeats / Q000235:genetics; D008954:Models, Biological; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D016133:Polymerase Chain Reaction; D013997:Time Factors; D014611:Vaccination
chemical_list: D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; C063317:L1 protein, Human papillomavirus type 16; D008156:Luciferases