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[Objective] To investigate the genomic phylogeny as well as the in vivo and in vitro infection and replication characteristics of the tick-borne parainfluenza virus type 5 (strain PIV5-JC12) recently isolated from Yunnan Province. [Methods] The strain PIV5-JC12 was identified through cytopathic effect (CPE) observation, negative staining electron microscopy, and an indirect immunofluorescence assay (IFA) based on the P protein. We determined its optimal culture temperature by comparing viral proliferation efficiency at 33 ℃ and 37 ℃ in Vero cells. A phylogenetic tree based on the whole genome was constructed via the maximum likelihood method to elucidate the molecular evolutionary characteristics of this strain. After infection of six representative cell lines of different origins (Vero, MDCK, HeLa, Huh7.5, MRC-5, and BHK-21), CPE observation, RT-qPCR, and tissue culture infectious dose 50% (TCID50) assays were employed to evaluate the virus replication kinetics. The in vivo pathogenicity of PIV5-JC12 was evaluated in Kunming (KM) and C57BL/6J mice. Mice were infected intranasally with high and low doses (2×107 TCID50 and 2×106 TCID50) and monitored for body weight and survival rate. The viral loads in various tissue and organ samples were measured at the time points of 1, 4, 7, and 10 days post-infection (dpi), and histopathological changes were examined. [Results] PIV5-JC12 induced CPEs, as manifested by the rounding and detachment of Vero cells. Irregular spherical particles with diameters of 50-200 nm were observed, and the specific P protein was identified by immunofluorescence. The tick-borne strain PIV5-JC12 showed the highest sequence identity with the tiger-borne strain PIV5 (OQ236149.1). Viral replication and proliferation in Vero cells were more efficient at 37 ℃ than at 33 ℃. PIV5-JC12 infected all the six cell lines of human, murine, simian, and canine origins. However, the CPEs varied among the cell lines, being more pronounced in Vero, MDCK and BHK-21 cells. Higher RNA replication efficiency was observed in Vero, MDCK, and HeLa cells. In vivo infection studies revealed differential susceptibility of KM and C57BL/6J mice to PIV5-JC12. The body weight loss (5%) was only observed in the high-dose infected C57BL/6J mice at 10 dpi. Lung viral loads in both mouse lines reached 104-105 copies/g, while no infectious virus was detected in the lung tissue at any of the time points examined. Histopathological staining analysis of lung tissue at the early stage (4 dpi) and late stage (10 dpi) of infection showed no significant pathological damage. [Conclusion] PIV5-JC12 isolated from tick samples from Yunnan Province shows broad cellular tropism and low pathogenicity in mice. The findings provide a basis for research on the pathogenic characteristics of PIV5 and its potential application as a vaccine vector.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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