A Highly Permissive K-C4 Cell Clone for Genotype II African Swine Fever Virus Reveals Passage-Dependent Hemadsorption Changes Associated with EP402R Deletion.
- 1Laboratory for Infectious Disease Prevention, Korea Zoonosis Research Institute, Jeonbuk National University, Iksan 54531, Republic of Korea.
- 2Department of Pharmacology, College of Pharmacy, Chonnam National University, Gwangju 61186, Republic of Korea.
- 3College of Veterinary Medicine and Bio-Safety Research Institute, Jeonbuk National University, Iksan 54596, Republic of Korea.
Abstract
African swine fever virus (ASFV) research commonly relies on primary porcine alveolar macrophages (PAMs), whose limited availability and variability hinder standardized virus isolation and propagation. This study aimed to establish a continuous cell line highly susceptible to genotype II ASFV and characterize phenotypic and genomic changes during serial passage. Thirty COS-1-derived clones were screened after infection with Korea/Pig/PAJU1/2019 using quantitative real-time polymerase chain reaction, immunofluorescence, and hemadsorption assays. Selected clones underwent serial passage, and viruses from passages 5 and 10 in the most susceptible clone were analyzed by whole-genome sequencing. K-C4 supported stable ASFV replication, with a mean viral genome level of 8.80 ± 0.55 log₁₀ copies/mL, compared with 7.38 ± 0.90 log₁₀ copies/mL in PAMs. Productive infection was confirmed by p72-specific immunofluorescence. The parental and passage 1 viruses retained hemadsorption, whereas the passage 5 virus lacked detectable hemadsorption despite continued replication, and hemadsorption reappeared by passage 10. Whole-genome analysis identified passage-associated deletions of differing extent within EP402R, encoding the hemadsorption-associated CD2-like viral protein (CD2v), which corresponded with the hemadsorption phenotype at each passage. These findings establish K-C4 as a highly permissive platform for genotype II ASFV propagation and characterization while emphasizing the need to monitor phenotypic and genomic stability during serial passage.
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BibTeX
@article{lee2026highly,
title = {{A Highly Permissive K-C4 Cell Clone for Genotype II African Swine Fever Virus Reveals Passage-Dependent Hemadsorption Changes Associated with EP402R Deletion.}},
author = {Eun-Gyeong Lee and Ju-Hun Kim and Sarah Lim and Sung-Hyun Moon and Hyunji Gu and Jun Young Park and Dae-Min Kim and Gun-Hee Lee and Ho-Seong Cho and Kyeong-Man Kim and Dongseob Tark},
journal = {Veterinary sciences},
year = {2026},
month = sep,
doi = {10.3390/vetsci13101024},
url = {https://europepmc.org/article/MED/42944120},
note = {CC BY},
}
RIS
TY - JOUR
TI - A Highly Permissive K-C4 Cell Clone for Genotype II African Swine Fever Virus Reveals Passage-Dependent Hemadsorption Changes Associated with EP402R Deletion.
AU - Eun-Gyeong Lee
AU - Ju-Hun Kim
AU - Sarah Lim
AU - Sung-Hyun Moon
AU - Hyunji Gu
AU - Jun Young Park
AU - Dae-Min Kim
AU - Gun-Hee Lee
AU - Ho-Seong Cho
AU - Kyeong-Man Kim
AU - Dongseob Tark
PY - 2026
DA - 2026/09/28
JO - Veterinary sciences
DO - 10.3390/vetsci13101024
UR - https://europepmc.org/article/MED/42944120
AB - African swine fever virus (ASFV) research commonly relies on primary porcine alveolar macrophages (PAMs), whose limited availability and variability hinder standardized virus isolation and propagation. This study aimed to establish a continuous cell line highly susceptible to genotype II ASFV and characterize phenotypic and genomic changes during serial passage. Thirty COS-1-derived clones were screened after infection with Korea/Pig/PAJU1/2019 using quantitative real-time polymerase chain reaction, immunofluorescence, and hemadsorption assays. Selected clones underwent serial passage, and viruses from passages 5 and 10 in the most susceptible clone were analyzed by whole-genome sequencing. K-C4 supported stable ASFV replication, with a mean viral genome level of 8.80 ± 0.55 log₁₀ copies/mL, compared with 7.38 ± 0.90 log₁₀ copies/mL in PAMs. Productive infection was confirmed by p72-specific immunofluorescence. The parental and passage 1 viruses retained hemadsorption, whereas the passage 5 virus lacked detectable hemadsorption despite continued replication, and hemadsorption reappeared by passage 10. Whole-genome analysis identified passage-associated deletions of differing extent within EP402R, encoding the hemadsorption-associated CD2-like viral protein (CD2v), which corresponded with the hemadsorption phenotype at each passage. These findings establish K-C4 as a highly permissive platform for genotype II ASFV propagation and characterization while emphasizing the need to monitor phenotypic and genomic stability during serial passage.
SN - 2306-7381
ER -