Fastidious human adenovirus type 40 can propagate efficiently and produce plaques on a human cell line, A549, derived from lung carcinoma.
Human adenovirus type 40 (Ad40) cannot propagate in conventional established human cell lines such as KB or HeLa cells. However, it has been shown that Ad40 DNA replicates in KB18 cells which express Ad2 E1B genes, suggesting that Ad40 is defective in the E1B gene function in KB or HeLa cells. We show here that Ad40 can propagate and produce plaques on A549 cells which do not contain Ad E1B genes. Our experiments show that the levels of replication of Ad40 DNA and production of infectious Ad40 virus in A549 cells are the same as or higher than those in 293 or KB18 cells. Dot blot analysis shows that the levels of Ad40 E1A and E1B mRNAs expressed in A549 cells at early to intermediate times postinfection are at least 10-fold higher than those in KB or KB18 cells. Northern (RNA) blot analysis shows that large E1B mRNA species (approximately 24S to 26S) are synthesized prior to the onset of DNA replication in A549 cells. No E1B mRNA species are synthesized in KB or KB18 cells at early times postinfection, and no differences in the expression of E1B mRNAs are seen between KB and KB18 cells. The experiment suggests that A549 cells have a cellular factor(s) which activates Ad40 E1B mRNA synthesis and that the E1B mRNA synthesis helps Ad40 propagation. In contrast, Ad40 can propagate in KB18 cells by using Ad2 E1B gene products that are constitutively expressed in this cell line. Furthermore, this result shows that Ad40 cannot propagate in KB cells because of the failure in the expression of E1B genes at early times postinfection.
Hashimoto
S
S
Meiji Institute of Health Science, Odawara, Japan.
Sakakibara
N
N
Kumai
H
H
Nakai
M
M
Sakuma
S
S
Chiba
S
S
Fujinaga
K
K
D017456:Adenovirus Early Proteins; D000260:Adenoviruses, Human / Q000254:growth & development* / Q000302:isolation & purification; D015152:Blotting, Northern; D003588:Cytopathogenic Effect, Viral; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis; D005243:Feces / Q000382:microbiology; D005814:Genes, Viral; D006801:Humans; D008175:Lung Neoplasms; D009856:Oncogene Proteins, Viral / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D012692:Serial Passage; D014407:Tumor Cells, Cultured; D010948:Viral Plaque Assay; D014776:Virus Cultivation
D017456:Adenovirus Early Proteins; D004279:DNA, Viral; D009856:Oncogene Proteins, Viral; D012367:RNA, Viral