Establishment and characterization of an immortalized renal cell line of the Chinese tree shrew (Tupaia belangeri chinesis)
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Authors: Gu, Tianle; Yu, Dandan; Li, Yu; Xu, Ling; Yao, Yu-Lin; Yao, Yong-Gang
Abstract
The Chinese tree shrew holds a great potential as a viable animal model in biomedical research, especially for infectious diseases and neuropsychiatric disorders. A thorough understanding of the innate immunity, which represents the first line that defends the host against viral infection, of the Chinese tree shrew, is needed. However, the progress is hindered by the lack of a proper cell line for research usage. In this study, we established a cell line that is applicable to the study of tree shrew innate immune responses against viral infections. The Chinese tree shrew primary renal cells (TSPRCs) were immortalized by simian virus 40 large T antigen (SV40LT) transduction, and the immortalized cells were termed TSR6 (tree shrew renal cell #6). TSR6 showed a similar morphology to TSPRCs and expressed the epithelial cell-specific marker cytokeratin 18 (KRT18). In addition, TSR6 could be transfected by transfection reagent and was suitable for CRISPR/Cas9-mediated gene editing. Infection of Newcastle disease virus (NDV) or herpes simplex virus 1 (HSV-1) in TSR6 induced the mRNA expression of tree shrew interferon- (tIFNB1) and myxovirus resistance protein 1 (tMx1) in a dose- and time-dependent manner. Collectively, we successfully established a tree shrew renal cell line and demonstrated that this cell line was suitable for the study of the innate immune response to viral infections.
Characterization and full genome sequencing of a velogenic Newcastle disease virus (NDV) strain Ck/IR/Beh/2011 belonging to subgenotype VII(L)
ACTA VIROLOGICA
Authors: Goudarzi, H.; Van Borm, S.; Bashashati, M.; Sabouri, F.; Abdoshah, M.; Nouri, A.; Banani, M.; Ebrahimi, M. M.; Molouki, A.
Abstract
Avian avulavirus 1, better known as Newcastle disease virus (NDV), causes substantial loss to the poultry industry in many developing countries. In this study we have characterized and fully sequenced the genome of a velogenic NDV strain named Beh (Ck/IR/Beh/2011) that has been used in our lab for a number of challenge and immunological studies over the last few years. This strain was isolated from poultry in the city of Behshahr, Mazandaran Province, Iran after an outbreak reported in the region in 2011. The intracerebral pathogenicity index (ICPI) was 1.8 in one-day-old chicks, characteristic of a velogenic NDV strain. Later, the virus was purified using a sucrose gradient centrifugation and used for next-generation sequencing (NGS). The results showed that the genome length was 15192 bp, similar to those of class II velogenic strains. In addition, the phylogenetic analysis based on the complete F gene showed that the NDV strain Beh has an F protein cleavage site (112)RRQKR down arrow F-117 and belongs to the newly identified subgenotype VII(L). Based on the biological and genetic characterization, NDV strain Beh is now the best documented reference isolate representing the novel subgenotype VII(L) in Iran.