HIV CYTOPATHICITY--ANTISENSE RNA APPROACH

Project: Research project

Description

The human immunodeficiency virus (HIV) exerts profound cytopathic
effects on the host T-lymphocyte following infection in vitro. The
cytopathogenic properties of HIV in vivo may directly account for
the depletion in circulating helper T4 lymphocytes and subsequent
immune dysfunction which is the underlying cause of the acquired
immune deficiency syndrome (AIDS). We hypothesize that
superinfection -- unrestricted virus replication and buildup of a
toxic viral gene product -- plays a crucial role in the process of
viral cytopathogenicity. The process of superinfection itself may
be directly influenced by biological properties of the host cell,
i.e., surface receptor expression such that cells low in this
receptor are not susceptible to virus mediated cytolysis. Thus,
reduced receptor expression may directly modulate superinfection
by preventing second round infection from progeny virus. We
hypothesize that regulating viral replication will interfere with
the process of viral superinfection, and the cytopathic effects of
virus infection may be abrogated. In the proposed study, antisense
viral RNAs will be used to confer cytopathicity resistance in
susceptible host cells by restricting virus replication and thus
prevent superinfection. Specifically, it is intended to: a)
evaluate whether cells expressing antisense viral RNA can be
protected from cytopathic effects of HIV infection. Inducible and
constitutive expression vectors, containing HIV genes in antisense
orientation will be introduced into HIV cytolysis susceptible cell
lines. The ability of these cells to resist cytopathic HIV
infection will be evaluated. To demonstrate the relationship
between viral cytopathicity and the buildup of toxic viral gene
products, the cytotoxic property of the envelope gene will be
determined by overexpressing this gene in susceptible cell lines.
b) investigate the dependence of viral cytopathogenicity and
superinfection on the level of T4 receptor expression on the host
cell, either by expressing the T4 gene in non-lymphoid cells in an
attempt to confer susceptibility to viral cytopathogenicity or
conferring cytolysis resistance in susceptible cells using
antisense T4 RNA expression vectors. c) evaluate ability of
retrovirus vectors expressing antisense HIV genes and synthetic
antisense HIV oligonucleotides to protect human lymphocytes from
cytopathic effect of HIv infection. Effectiveness of antisense RNA
inhibition in restricting viral replication and cytopathicity in
vitro may provide the rationale for the use of such an approach in
vivo.
StatusFinished
Effective start/end date12/1/8811/30/93

Funding

  • National Institutes of Health
  • National Institutes of Health: $94,991.00
  • National Institutes of Health: $103,287.00
  • National Institutes of Health
  • National Institutes of Health

Fingerprint

Antisense RNA
HIV
Superinfection
CD4-Positive T-Lymphocytes
Genes
Virus Diseases
Virus Replication
Viral Cytopathogenic Effect
Infection
RNA
Thyroid Hormone Receptors
Poisons
Viral RNA
Cell Surface Receptors
Viral Proteins
Oligonucleotides
Lymphocytes
Viruses
T-Lymphocytes
Cell Line

ASJC

  • Medicine(all)
  • Immunology and Microbiology(all)