A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo.
A deletion mutant of severe acute respiratory syndrome coronavirus (SARS-CoV) has been engineered by deleting the structural E gene in an infectious cDNA clone that was constructed as a bacterial artificial chromosome (BAC). The recombinant virus lacking the E gene (rSARS-CoV-DeltaE) was rescued in Vero E6 cells. The recovered deletion mutant grew in Vero E6, Huh-7, and CaCo-2 cells to titers 20-, 200-, and 200-fold lower than the recombinant wild-type virus, respectively, indicating that although the E protein has an effect on growth, it is not essential for virus replication. No differences in virion stability under a wide range of pH and temperature were detected between the deletion mutant and recombinant wild-type viruses. Although both viruses showed the same morphology by electron microscopy, the process of morphogenesis seemed to be less efficient with the defective virus than with the recombinant wild-type one. The rSARS-CoV-DeltaE virus replicated to titers 100- to 1,000-fold lower than the recombinant wild-type virus in the upper and lower respiratory tract of hamsters, and the lower viral load was accompanied by less inflammation in the lungs of hamsters infected with rSARS-CoV-DeltaE virus than with the recombinant wild-type virus. Therefore, the SARS-CoV that lacks the E gene is attenuated in hamsters, might be a safer research tool, and may be a good candidate for the development of a live attenuated SARS-CoV vaccine.
DeDiego
Marta L
ML
Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Darwin 3, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.
Alvarez
Enrique
E
Almazán
Fernando
F
Rejas
María Teresa
MT
Lamirande
Elaine
E
Roberts
Anjeanette
A
Shieh
Wun-Ju
WJ
Zaki
Sherif R
SR
Subbarao
Kanta
K
Enjuanes
Luis
L
D000818:Animals; D018938:Caco-2 Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D017353:Gene Deletion; D006801:Humans; D012137:Respiratory System / Q000821:virology; D045473:Severe acute respiratory syndrome-related coronavirus / Q000502:physiology*; D045169:Severe Acute Respiratory Syndrome / Q000821:virology*; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000235:genetics*; D014779:Virus Replication
D015678:Viral Structural Proteins