Important note: Information in this article was accurate in 1988. The state of the art may have changed since the publication date.
CONSTITUTIVE PRODUCTION OF A REVERSE-TRANSCRIPTASE-NEGATIVE AIDS RETROVIRUS PARTICLE
Vaccines 87. Modern Approaches to New Vaccines: Prevention of AIDS and Other Viral, Bacterial, and Parasitic Diseases. Chanock RM et al, eds. New York, Cold Spring Harbor Laboratory, p. 143-5, 1987.. Unique Identifier : AIDSLINE ICDB/88647968 Folks TM; Justement JS; Powell D; Kinter A; Rabson A; Fauci AS; Lab. of Immunoregulation, NIAID, NIH, Bethesda, MD 20892
Abstract:
A cell line (8E5) was cloned containing a defective proviral genome that constitutively produces acquired immune deficiency syndrome (AIDS) viral particles. When culture supernatants from the 8E5 cell line were collected and tested for the presence of AIDS viral antigens by an antigen-capture method, it was found that equivalent amounts of viral antigen were produced by the 8E5 cells when compared with cells infected with wild-type virus. When culture supernatants from 8E5 cell line were added to A3.01 cells, which are sensitive to the cytopathic effects of the AIDS retrovirus, no evidence of cytopathic effect (CPE) or viral replication could be detected. These results indicated that a critical mutation had occurred, which permitted viral proteins to be produced but restricted the production to noninfectious particles only. Southern, Northern, and Western blots were performed on DNA, RNA, and proteins, respectively, from the 8E5 cell line. All of the major viral proteins (gP120, P55, gP41, and P25) were present in the 8E5 cell line except P64 (reverse transcriptase) and P31 (endonuclease). This finding indicates that the 8E5 cell line produces most of the major structural proteins of human immunodeficiency virus (HIV). The 8E5 cell line does not produce the reverse transcriptase and endonuclease proteins since its pol gene contains a premature termination codon. The viral particles derived from this cell line could be used as an attenuated virus in nonhuman primates and could serve to delineate the nature and scope of protective immunity against wild-type viral infections, which could then be extrapolated to humans. In addition, the strategy of generating clones that produce defective viral particles may ultimately lead to the development of new mutant viruses for vaccine purposes. (4 Refs)
Keywords: Cell Line Cloning, Molecular Cytopathogenic Effect, Viral Defective Viruses/*GENETICS Human HIV/ENZYMOLOGY/*GENETICS/PHYSIOLOGY Mutation RNA-Directed DNA Polymerase/*GENETICS *Virus Replication MEETING PAPER
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Important note: Information in this article was accurate in 1988. The state of the art may have changed since the publication date.
