Research Article: Screening of Potential HIV-1 Inhibitors/Replication Blockers Using Secure Lentiviral in Vitro System

Date Published: , 2011

Publisher: A.I. Gordeyev

Author(s): M.M. Prokofjeva, P.V. Spirin, D.V. Yanvarev, A.V. Ivanov, M.S. Novikov, O.A. Stepanov, M.B. Gottikh, S.N. Kochetkov, B. Fehse, C. Stocking, V.S. Prassolov.

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Abstract

The development and usage of safe cell systems for testing agents which possess
anti-HIV activity is a very important factor in the design of new drugs. We have
described in detail a system we designed that is based on lentiviral vectors
(Prokofjeva et. al.,Antiviral Therapy,in print) for swift and
completely safe screening of potential HIV-1 replication inhibitors. The system
enables one to test the efficiency of the inhibitory activity of compounds whose
action is directed towards either wild-type HIV-1 reverse transcriptase or
integrase, or mutant enzymes corresponding to the drug-resistant virus form.
Testing results of a number of already known drugs, which correlate well with
published data as well as data on newly synthesized compounds, were obtained.
Application of this system substantially broadens the possibilities of
preclinical anti-HIV drugs testing.

Partial Text

The human immunodeficiency virus type 1 (HIV-1), which belongs to the lentivirus
genus of the retrovirus family, is responsible for one of the most common and
life-threatening diseases known as the acquired immunodeficiency syndrome (AIDS).
According to the World Health Organization (WHO), by the end of 2008, the number of
HIV-1-infected people topped 33 million [1].
In 2010, official records put the number of HIV-1-infected people in Russia at
520,000  [2]. It should be noted that in
reality the actual number of infected people can be two or even three times higher.
It follows from the prognoses of the WHO and non-governmental organizations that
even if all the initiatives to control AIDS propagation were implemented and
anti-HIV therapy was used, the number of HIV-infected people may still exceed
48 million in the next several years.

Cell cultivation

Construction of pseudo-HIV-1 particles and using them to infect different
eukaryotic cell lines

A number of human and mouse cell lines were used to demonstrate that the described
system for safe screening of potential HIV-1 replication inhibitors allows one to
test the inhibitory activity of the compounds, whose action is directed both towards
the reverse transcriptase and integrase of wild-type HIV-1 and towards their mutant
forms corresponding to drug-resistant forms of the virus. It is important that the
pseudo-HIV-1 particles used in this system are noninfectious. They are actually
single-acting viruses (recombinant lentiviral vectors) that contain a complete set
of viral enzymes ensuring the synthesis of the recombinant two-stranded DNA provirus
and its integration into the genome of target cells. Next, the cell systems allow
the expression of marker genes, which were integrated into the cell genome, within
the recombinant genome of pseudo-HIV-1 particles.

 

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