ABSTRACT
A comparative analysis of reproduction of 9 strains of hepatitis A viruses (HSA-15, CF-979, MI, MBB 11/5, H-141, KMW-1, GBM, IH-26, WR-61) from different regions of the world in cell cultures (PLC/PRF/5, HEL-240, FRhK-4, MK) revealed the differences in the capacity of the viruses for reproduction in these cell lines. The factors influencing HAV reproduction in cell cultures such as temperature, medium, and sera were studied. In one-cycle infection, accumulation of vRNA and formation of virus particles were analysed.
Subject(s)
Hepatovirus/physiology , Virus Replication/physiology , Animals , Cell Line , Cells, Cultured/microbiology , Culture Media , Hepatovirus/analysis , Hepatovirus/isolation & purification , Humans , RNA, Viral/analysis , Temperature , Virion/physiology , Virus Cultivation/methodsABSTRACT
A HAV strain derived from a patient in Moscow multiplied in a continuous cell line PLC/PRF/5 at 32 degrees C (variant MI-1) and at 37 degrees C (variant MI-1.1). These two variants of the strain MI retained the ability for reproduction both at 32 degrees C and 37 degrees C after passages in cell culture. The strain MI also replicates in MK cells. Negative results on HAV cultivation were obtained in HEF-240 and FRhK-4 cell cultures. The MI-1 and MI-1.1 variants are typical of hepatitis A viruses by their physicochemical properties, the results of ELISA, protein electrophoresis and dot hybridization tests.
Subject(s)
Adaptation, Physiological/physiology , Hepatovirus/physiology , Virus Replication/physiology , Animals , Cell Line , Cells, Cultured/microbiology , Chlorocebus aethiops , DNA, Viral/genetics , Electrophoresis, Polyacrylamide Gel , Embryo, Mammalian , Hepatitis A/microbiology , Hepatovirus/isolation & purification , Hepatovirus/ultrastructure , Humans , Macaca mulatta , Microscopy, Electron , Nucleic Acid Hybridization , Viral Proteins/analysis , Virus Cultivation/methodsABSTRACT
Using the MBB/11/5 strain originally adapted to the PLC/PRF/5 cell line, reproduction of the virus in diploid cells of human embryo fibroblasts, continuous primate lines (RAMT, FRhK-4) and human urinary bladder tumor (T-24) lines was studied. We obtained HAV DNA sequences (about 99% from vRNA) for the MBB/11/5 strain which were used as a probe for demonstration of vRNA synthesis in the infected cells.
